GSM8878376: C3.clean; Mus musculus; RNA-Seq
Total RNA was extracted from CD4+ T cells by using TRIzol Reagent (Invitrogen, Cat NO. 15596026). The RNA purity, concentration, and integrity were evaluated with an Agilent TapeStation 4200 (Agilent Technologies). RNA sequencing library was prepared by mRNA Library Prep Kit for Illumina®, and the indexed libraries were subjected to sequencing on DNBSEQ-T7 sequencer with PE150 model. Read mapping was performed via STRA software (version 2.5.3a) using CHOK1GS from Ensembl as a reference genome. Afterwards, high-quality reads mapped to the exon regions of each gene were counted by featureCounts (Subread-1.5.1; Bioconductor) and then RPKMs were calculated. Differential expression analysis was identified using the edgeR package (version 3.12.1). 2 μg total RNAs were used for stranded RNA sequencing library preparation using KCTM Stranded mRNA Library Prep Kit for Illumina® (Catalog NO. DR08402, Wuhan Seqhealth Co., Ltd. China) following the manufacturer's instruction. PCR products corresponding to 200-500 bps were enriched, quantified and finally sequenced on DNBSEQ-T7 sequencer (MGI Tech Co., Ltd. China) with PE150 model.