Hepatocyte and Leukemia Cell Culture Protocols

Human hepatoma cells (HepG2) were maintained in DMEM supplemented with 10% fetal bovine serum (FBS, Hyclone). Human T-cell leukemia cells (Jurkat) were maintained in RPMI-1640 supplemented with 10% FBS. Primary hepatocytes were isolated and cultured as previously described (Fan et al., 2020 (link)). Mouse Npnt promoter-luciferase constructs (Lanthier et al., 2011 (link)) and BRG1 expression constructs have been previously described (Chen et al., 2020c (link); Li et al., 2020a,b (link), c (link)). Small interfering RNAs were purchased from Dharmacon. Transient transfections were performed with Lipofectamine 2000. Luciferase activities were assayed 24–48 h after transfection using a luciferase reporter assay system (Promega) as previously described (Wu et al., 2020 (link); Yang et al., 2020a,b (link)). For conditioned media (CM) collection, the cells were switched to and incubated with serum-free media overnight. The next day, the media were collected, centrifuged at 4000 × g for 30 min at 4°C using 3-kDa MW cut-off filter units (Millipore) and sterilized through a 0.4-μm filter.