SRX3407594 GSM2861485 SRP125260 SRS2706608 GSM2861485 miRNA-Seq TRANSCRIPTOMIC size fractionation Then the infected Vero cell monolayers were harvested, thereafter, the isolation of the parasites from host cells were performed by passaging through a 20 G needle one time and a 27 G needle three times. Cell debris was removed and parasites were purified by 40% percoll gradient centrifugation. Purified parasites were washed two times with fresh DMEM by centrifuging at 1000×g for 10 min. After washing, the pellet was resuspended in fresh medium and the number of tachyzoites was counted with a hemocytometer counting chamber. Total RNAs of N. caninum were extracted using Trizol reagent (Invitrogen, SF, USA) performed as instructed in protocol provided by manufacturer. The quantification and quality of RNA purification were evaluated by Nanodrop 2000 spectrophotometer (Thermo Scientific CA, USA). Standard agarose gel electrophoresis was carried out to analyze the integrity of isolated total RNA. The purified total RNA was stored at -80ºC until cDNA library construction. Illumina HiSeq 2000 gds 302861485 GSM2861485 GEO Accession GSM2861485 SRX3407595 GSM2861486 SRP125260 SRS2706609 GSM2861486 miRNA-Seq TRANSCRIPTOMIC size fractionation Then the infected Vero cell monolayers were harvested, thereafter, the isolation of the parasites from host cells were performed by passaging through a 20 G needle one time and a 27 G needle three times. Cell debris was removed and parasites were purified by 40% percoll gradient centrifugation. Purified parasites were washed two times with fresh DMEM by centrifuging at 1000×g for 10 min. After washing, the pellet was resuspended in fresh medium and the number of tachyzoites was counted with a hemocytometer counting chamber. Total RNAs of N. caninum were extracted using Trizol reagent (Invitrogen, SF, USA) performed as instructed in protocol provided by manufacturer. The quantification and quality of RNA purification were evaluated by Nanodrop 2000 spectrophotometer (Thermo Scientific CA, USA). Standard agarose gel electrophoresis was carried out to analyze the integrity of isolated total RNA. The purified total RNA was stored at -80ºC until cDNA library construction. Illumina HiSeq 2000 gds 302861486 GSM2861486 GEO Accession GSM2861486