GSM7679576: NK cells rep2; Rattus norvegicus; RNA-Seq
Cell sorting was performed with a FACSAria III (BD Biosciences, NJ, USA). CD4 T cells (CD45+/CD11b-/CD3+/CD4+), CD8 T cells (CD45+/CD11b-/CD3+/CD8a+), NK cells (CD45+/CD11b-/CD3-/CD161+), and B cells (CD45+/CD11b-/CD3-/CD45RA+) were obtained from the rat spleen. Cells were suspended in wash buffer containing 5% 7-AAD solution (559925, BD Pharmgen) 5 min before analysis to exclude dead cells. The sorted cells were at once pooled in RPMI-1640 containing 10% fetal bovine serum. After centrifugation at 400 g at 4 °C for 5 min, total RNA was prepared using ISOGEN II (311-07361, Nippon Gene Co., Tokyo, Japan) and purified using an RNeasy Plus Mini Kit (74136, Qiagen, Limburg, Netherland) with gDNA elimination using an RNase-Free DNase set (79254, Qiagen) according to the manufacturer's protocols. Total RNA was used to prepare RNA-Seq libraries with a NEBNext Ultra ll Directional RNA Library Prep Kit for Illumina.