GSM6001228: T-bet ZsGreen Low rep2 (RNA-seq); Mus musculus; RNA-Seq
For each sort, spleens and lymph nodes of 4-5 mice were pooled. Thy1.1+ CD4 T cells were magnetically enriched with CD90.1 beads. Live, single CD90.1+ lymphocytes were FACS sorted according to their T-bet ZsGreen reporter brightness into high and low expressors. For RNA-Seq: Cells were lysed using RA1 buffer and stored at -80°C until further processing. Total RNA was purified with Nucleospin RNA XS Micro Kit without carrier RNA. For ATAC-Seq: The cell sample preparation manual from the ATAC-Seq Kit (activemotif) was followed for DNA tagmentation and purification. The samples were stored at -20°C until further processing. For RNA-Seq: cDNA library was prepared using Smart-Seq v4 mRNA Ultra Low Input RNA Kit. For ATAC-Seq: PCR Amplification of tagmented DNA was performed using the ATAC-Sequencing Kit (activemotif). With an Illumina NextSeq2000 device, paired-end sequencing (2x75bp) of the libraries was performed.