title: GSM8371781: TNFi-treated, replicate 4, snRNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Briefly, samples were cut into approximately 7-mm pieces, injected with 1 ml of ice-cold Nuclei EZ Lysis buffer (NUC101; MilliporeSigma) supplemented with a protease (589279100; Hoffmann-La Roche Ltd, Basel, Switzerland) and ribonuclease (RNAse, N2615; Promega Corp., Madison, WI, USA) inhibitor (250 units/ml RNasin® Plus N2615; Promega Corp., Madison, WI, USA) using a 26-gauge needle, minced into 1 to 2-mm pieces with scissors in a weigh boat with 1 ml additional supplemented lysis buffer, and then transferred to a gentleMACSTM C tube (Miltenyi Biotec, Bergisch Gladbach, Germany). The gentleMACS lung1 and lung 2 programs were run in sequence, with the latter stopped after 20 seconds. The foam was spun down for 10 seconds using a short spin. The suspension was passed through a 40-µm cell strainer and washed with 4℃ cold PBS containing 1% bovine serum albumin (BSA). Nuclei were pelleted at 500×g and resuspended in 1X PBS containing 1% BSA and 0.5 units/µl RNasin Plus before counting them by hemocytometer and diluting to 1,000 nuclei/µl. 10x Chromium libraries were prepared according to the manufacturer's protocol snRNA-seq