SRX18332037 GSM6745905_r1 SRP409011 PRJNA903907 SRS15821102 GSM6745905 GSM6745905 RNA-Seq TRANSCRIPTOMIC cDNA Testes from from wild-type and Dhx36 mutant spermatogenesis synchronous mice were collected in PBS on ice. After removal of the tunica albuginea, the testes were incubated in 5mL of PBS with 120 U/mL of collagenase type I (Worthington) at 32 °C with gentle agitation for 5 min. The dispersed seminiferous tubules were then digested with 5mL of 0.25% trypsin (Gibco), 0.1 mL of DNase I (5 mg/mL; Sigma) at 32 °C for 8 min, and then terminated by adding 0.5 mL of fetal bovine serum (FBS; Gibco) to inactivate trypsin. The resulting dissociated testicular cells were filtered through a PBS-prewetted cellular filter (70 μm). After spin down at 500 g for 5 min at 4 °C, the cells in the pellet were resuspended at a concentration of 106 cells/mL in DMEM with Hoechst 33342 (3 mg/mL; Sigma) and 5 μl DNase I followed by rotating for 20 min at 32 °C in the oven at 10 r.p.m./min speed. Zygotene spermatocytes were collected based on their fluorescent label with Hoechst 33342 staining using FACS (BD Biosciences). Cell purity was further examined by meiotic nuclear spreading with immunofluorescence staining as described above. The purity of isolated zygotene is about 95% Total RNA was extracted from zygotene spermatocytes isolated from wild-type and Dhx36 mutant spermatogenesis synchronous mice using Trizol reagent (Invitrogen). RNA quality was determined by Qubit Fluorometer and Agilent Bioanalyzer 2100. rRNAs were removed from total RNAs by the NEBNext rRNA Depletion Kit (NEB). The remaining RNAs were fragmented and then reverse transcribed. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions. Illumina HiSeq 2000 SRX18332038 GSM6745906_r1 SRP409011 PRJNA903907 SRS15821103 GSM6745906 GSM6745906 RNA-Seq TRANSCRIPTOMIC cDNA Testes from from wild-type and Dhx36 mutant spermatogenesis synchronous mice were collected in PBS on ice. After removal of the tunica albuginea, the testes were incubated in 5mL of PBS with 120 U/mL of collagenase type I (Worthington) at 32 °C with gentle agitation for 5 min. The dispersed seminiferous tubules were then digested with 5mL of 0.25% trypsin (Gibco), 0.1 mL of DNase I (5 mg/mL; Sigma) at 32 °C for 8 min, and then terminated by adding 0.5 mL of fetal bovine serum (FBS; Gibco) to inactivate trypsin. The resulting dissociated testicular cells were filtered through a PBS-prewetted cellular filter (70 μm). After spin down at 500 g for 5 min at 4 °C, the cells in the pellet were resuspended at a concentration of 106 cells/mL in DMEM with Hoechst 33342 (3 mg/mL; Sigma) and 5 μl DNase I followed by rotating for 20 min at 32 °C in the oven at 10 r.p.m./min speed. Zygotene spermatocytes were collected based on their fluorescent label with Hoechst 33342 staining using FACS (BD Biosciences). Cell purity was further examined by meiotic nuclear spreading with immunofluorescence staining as described above. The purity of isolated zygotene is about 95% Total RNA was extracted from zygotene spermatocytes isolated from wild-type and Dhx36 mutant spermatogenesis synchronous mice using Trizol reagent (Invitrogen). RNA quality was determined by Qubit Fluorometer and Agilent Bioanalyzer 2100. rRNAs were removed from total RNAs by the NEBNext rRNA Depletion Kit (NEB). The remaining RNAs were fragmented and then reverse transcribed. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions. Illumina HiSeq 2000 SRX18332039 GSM6745907_r1 SRP409011 PRJNA903907 SRS15821104 GSM6745907 GSM6745907 RNA-Seq TRANSCRIPTOMIC cDNA Testes from from wild-type and Dhx36 mutant spermatogenesis synchronous mice were collected in PBS on ice. After removal of the tunica albuginea, the testes were incubated in 5mL of PBS with 120 U/mL of collagenase type I (Worthington) at 32 °C with gentle agitation for 5 min. The dispersed seminiferous tubules were then digested with 5mL of 0.25% trypsin (Gibco), 0.1 mL of DNase I (5 mg/mL; Sigma) at 32 °C for 8 min, and then terminated by adding 0.5 mL of fetal bovine serum (FBS; Gibco) to inactivate trypsin. The resulting dissociated testicular cells were filtered through a PBS-prewetted cellular filter (70 μm). After spin down at 500 g for 5 min at 4 °C, the cells in the pellet were resuspended at a concentration of 106 cells/mL in DMEM with Hoechst 33342 (3 mg/mL; Sigma) and 5 μl DNase I followed by rotating for 20 min at 32 °C in the oven at 10 r.p.m./min speed. Zygotene spermatocytes were collected based on their fluorescent label with Hoechst 33342 staining using FACS (BD Biosciences). Cell purity was further examined by meiotic nuclear spreading with immunofluorescence staining as described above. The purity of isolated zygotene is about 95% Total RNA was extracted from zygotene spermatocytes isolated from wild-type and Dhx36 mutant spermatogenesis synchronous mice using Trizol reagent (Invitrogen). RNA quality was determined by Qubit Fluorometer and Agilent Bioanalyzer 2100. rRNAs were removed from total RNAs by the NEBNext rRNA Depletion Kit (NEB). The remaining RNAs were fragmented and then reverse transcribed. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions. Illumina HiSeq 2000 SRX18332040 GSM6745908_r1 SRP409011 PRJNA903907 SRS15821105 GSM6745908 GSM6745908 RNA-Seq TRANSCRIPTOMIC cDNA Testes from from wild-type and Dhx36 mutant spermatogenesis synchronous mice were collected in PBS on ice. After removal of the tunica albuginea, the testes were incubated in 5mL of PBS with 120 U/mL of collagenase type I (Worthington) at 32 °C with gentle agitation for 5 min. The dispersed seminiferous tubules were then digested with 5mL of 0.25% trypsin (Gibco), 0.1 mL of DNase I (5 mg/mL; Sigma) at 32 °C for 8 min, and then terminated by adding 0.5 mL of fetal bovine serum (FBS; Gibco) to inactivate trypsin. The resulting dissociated testicular cells were filtered through a PBS-prewetted cellular filter (70 μm). After spin down at 500 g for 5 min at 4 °C, the cells in the pellet were resuspended at a concentration of 106 cells/mL in DMEM with Hoechst 33342 (3 mg/mL; Sigma) and 5 μl DNase I followed by rotating for 20 min at 32 °C in the oven at 10 r.p.m./min speed. Zygotene spermatocytes were collected based on their fluorescent label with Hoechst 33342 staining using FACS (BD Biosciences). Cell purity was further examined by meiotic nuclear spreading with immunofluorescence staining as described above. The purity of isolated zygotene is about 95% Total RNA was extracted from zygotene spermatocytes isolated from wild-type and Dhx36 mutant spermatogenesis synchronous mice using Trizol reagent (Invitrogen). RNA quality was determined by Qubit Fluorometer and Agilent Bioanalyzer 2100. rRNAs were removed from total RNAs by the NEBNext rRNA Depletion Kit (NEB). The remaining RNAs were fragmented and then reverse transcribed. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions. Illumina HiSeq 2000