GSM9407384: HLMC_D4_Control [S4]; Homo sapiens; RNA-Seq
Human lung tissue was cut into small pieces and enzymatically digested for 30 minutes with 0.2 mg/mL DNase I (Sigma-Aldrich, St. Louis, MO, USA) and 0.1 mg/mL Liberase TL (Sigma-Aldrich). Thereafter, the tissue was vigorously cut by scissors and mechanically disrupted by plunging through a syringe. The cells were purified by 30% Percoll centrifugation, and red blood cells were lysed using ACK lysis buffer (Thermo Fisher, Waltham, MA, USA). Mast cells were enriched by magnetic-activated cell sorting (MACS) enrichment using CD117 MicroBead Kit (Miltenyi). RNA was extracted using Arcturus PicoPure RNA Isolation Kit (Thermo Fisher Scientific). The libraries were prepared at NGI Science for Life Laboratory, Stockholm, Sweden using SMARTer Total Stranded RNA-seq (Takara Bio).