GSM6691978: CH4; Mus musculus; RNA-Seq
isolation protocol: Succesive CD4+ and C62L+ T cell isolation using positive selection with MACS Beads from wildtype mouse splenocytes. Cells were either harvested directly, or activated for 48 hrs with anti-CD3 and anti-CD28 under Th1, Th2 or Th17 polarizing conditions and the cultured for 1 week or repeatedly restimulated with anti-CD3 and anti-CD28 and cultured for 4 weeks. After each week the viable cells were collected by Ficoll grandient centrifugation. FACS sort: none, viable cells were obtained by Ficoll gradient centrifugation RNA isolation protocol: RNA of cells was extracted using RNeasy Plus Micro Kit (Qiagen) and RNA quality was assessed by an Agilent 2100 Bioanalyzer. Illumina® Stranded Total RNA Prep, Ligation with Ribo-Zero Plus