GSM8459920: PBS 24 hours; Mus musculus; RNA-Seq
Livers were harvested in RPMI supplemented with 10% FCS and digested with collagenase IV (1 mg/ml, Thermo Fisher Scientific) and DNase I (50 µg/ml, Roche) for 45 minutes at 37°C on a rotating device. Cells were washed with PBS containing 2 mM EDTA and filtered through a 70-µm strainer (Biologix). Red blood cells were lysed using a red blood cell lysis buffer (17 mM Tris pH 7.2, 144 mM NH4Cl) for 2 minutes and cells were washed with PBS. Lymphocyte enrichment was performed using a density gradient centrifugation step (Percoll, Sigma-Aldrich). Library amplification was carried out following the BD Rhapsody System mRNA Whole Transcriptome Analysis (WTA) and Sample Tag Library Preparation Protocol (Doc ID: 23-24119 (02))