Fig. 5
From: DNA damage sensitivity of SWI/SNF-deficient cells depends on TFIIH subunit p62/GTF2H1
Cancer cells with chronic BRG1 deficiency restore GTF2H1 expression. a Immunoblot showing total protein levels of BRM, BRG1, and GTF2H1, in cell lysates of U2OS and BRG1-deficient non-small lung cancer cell (NSCLC) lines A549 and H1299 treated with control (CTRL), BRG1 or BRM siRNAs. Ku70 was used as loading control. b Relative quantification of GTF2H1 protein levels in U2OS, A549, and H1299 cells transfected with control (CTRL), BRG1 or BRM siRNA. GTF2H1 levels were normalized to Ku70 and the total relative amount of GTF2H1 in whole cell lysates was set to 1.0 in U2OS siCTRL. Mean & S.E.M. from at least three independent experiments **P < 0.01, ***P < 0.001, n.s., non-significant. c A549 cells with and without stable expression of GFP-GTF2H1, driven by the ectopic PGK promoter, were treated with control (CTRL), BRM or BRG1 siRNAs. Cell lysates were analyzed by immunoblotting against BRM and GTF2H1. Ku70 was used as loading control. d A549 cells, with or without stable expression of GFP-GTF2H1 were seeded 48 h after transfection with control (CTRL), BRM or BRG1 siRNAs, in triplicate, at a density of 1000 cells per well and grown for 12 before fixation and staining. e Quantification of colony forming capacity of A549 (shown in d) and H1299 (shown in Supplementary Fig. 4e) cell lines with or without stable GFP-GTF2H1 expression and treated with control (CTRL), BRM or BRG1 siRNAs. Clonal capacity was normalized to 100% in control conditions (CTRL). Mean & S.E.M. of three independent experiments, each performed in triplicate. ***P < 0.001, n.s., non-significant, relative to siCTRL. Full-size immunoblot scans are provided in Supplementary Fig. 7a,b
