ERX4146937 E-MTAB-9144:D5_0.05_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597467 SAMEA6869899 D5_0.05_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 5 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146938 E-MTAB-9144:D19_0.05_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597468 SAMEA6869900 D19_0.05_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 19 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146939 E-MTAB-9144:D9_0.05_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597469 SAMEA6869901 D9_0.05_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 9 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146940 E-MTAB-9144:D19_0.5_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597470 SAMEA6869902 D19_0.5_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 19 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146941 E-MTAB-9144:D5_0.5_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597471 SAMEA6869903 D5_0.5_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 5 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146942 E-MTAB-9144:D9_0.5_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597472 SAMEA6869904 D9_0.5_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 9 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 0.5 ERX4146943 E-MTAB-9144:D5_6_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597473 SAMEA6869905 D5_6_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 5 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 6 ERX4146944 E-MTAB-9144:D19_6_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597474 SAMEA6869906 D19_6_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 19 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 6 ERX4146945 E-MTAB-9144:D9_6_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597475 SAMEA6869907 D9_6_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 9 Experimental Factor: compound polystyrene microplastics Experimental Factor: dose 6 ERX4146946 E-MTAB-9144:D19_C_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597476 SAMEA6869908 D19_C_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 19 Experimental Factor: compound none ERX4146947 E-MTAB-9144:D5_C_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597477 SAMEA6869909 D5_C_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 5 Experimental Factor: compound none ERX4146948 E-MTAB-9144:D9_C_s ERP122049 PRJEB38614 RNA-Seq of Oryzias melastigma treated with polystyrene microplastics against untreated controls ERS4597478 SAMEA6869910 D9_C_s RNA-Seq TRANSCRIPTOMIC RT-PCR The embryos were collected within 3-5 h after the fish started to lay eggs and then transferred into glass Petri dishes. Three different sizes of polystyrene microplastics with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of polystyrene microplastic solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; the concentrations used in this study are according to the environmentally relevant concentrations and previous literature on the toxicity of MPs on aquatic organisms. The controls were exposed to an equal volume of artificial seawater (30‰ salinity) alone. 150 healthy embryos were selected randomly and transferred into glass Petri dishes with 100 mL solutions. All the treatments were performed in triplicate. Two-thirds of the test solution in each dish was replaced daily with the same concentration of MPs during the experimental period. The newly hatched larvae were collected daily and placed in new dishes, and the PS MP solution or seawater was renewed every 24 h. During the exposure period, dead embryos were taken out immediately and recorded. The pooled samples of 15 embryos on 3, 6, or 9 dpf or the whole larva on 19 dpf from each duplicate were frozen immediately in liquid nitrogen and stored at -80ºC to determine the mRNA expression levels via RNA sequencing (RNA-Seq) and target gene expression profiles via real-time quantitative PCR (RT-qPCR). The larval fish were exposed for 19 dpf and then transferred to clear seawater. Fish were maintained in the laboratory in aerated 30‰ artificial seawater at 6.0 ± 0.2 mg O2 L−1 at 28 ± 2ºC in a 14-h light:10-h dark cycle. The adult fish (4-month old) were fed with newly hatched brine shrimp (Artemia salina) nauplii twice daily at 2% body weight. Three different sizes of PS MPs with diameters of 0.05, 0.50, and 6.00 μm were used, and four concentrations of PS MP solutions were used as treatments: 0 (control group), 0.1, 1×103 and 1×106 particles/mL for each diameter of MPs; Total RNA was harvested using TRIZOL reagent. The sample for transcriptome sequencing consisted of equal numbers of total RNA samples at 5, 9 and 19 dpf. Construction of cDNA libraries was performed according to the manufacturer's instructions (Illumina, USA). The library products were ready for sequencing using the BGISEQ-500 Sequencing platform (BGI, China), and the sequencing results were de novo assembled using Trinity. Illumina HiSeq 2000 Experimental Factor: time 9 Experimental Factor: compound none