Fig. 6

DNA damage sensitivity of BRM-deficient cells correlates with GTF2H1 expression. a Immunoblot of BRM, BRG1, and GTF2H1 in MRC5 wild-type (WT) cells and cells transfected with sgRNA against BRM or BRG1. b Quantification of GTF2H1 levels in immunoblot shown in a, corrected by tubulin loading control, and set to 1.0 in MRC5 WT. c IF of total GTF2H1 and BRM levels in MRC5 WT and sgBRM knockout clones c1, c3, c6, and c7. Scale bar: 10 μm. d Quantification of GTF2H1 IF signal (shown in c). Total GTF2H1 levels were normalized to MRC5 WT, set to 1.0. Mean & S.E.M. of > 200 cells from two independent experiments. e Immunoblot of BRM, BRG1, and GTF2H1 levels in MRC5 WT and sgBRM clones c1, c3, c6, and c7. f Quantification of GTF2H1 levels shown in e, as described in b, using Ku70 as loading control. Mean & S.E.M. from four independent experiments. g GTF2H1 levels in a mixed population of MRC5 sgBRM knockout clone c1 cells non-transfected or transfected with BRM-GFP or BRG1-GFP. Scale bar: 5 μm. h XPD recruitment to LUD in MRC5 WT and sgBRM knockout clones c1 and c3, 30 min after damage. Scale bar: 5 μm. i Relative quantification of XPD recruitment to LUD (shown in h) in MRC5 WT and BRM knockout clones c1 and c3, normalized to WT, as described in the methods. Mean & S.E.M. of > 65 cells per sample. j UV-C colony survival of MRC5 WT cells and BRM knockout clones c1 and c3. k Cisplatin colony survival of MRC5 WT and BRM knockout clones c1 and c3. l UV-C colony survival of MRC5 WT cells and BRM knockout clones c6 and c7. m Cisplatin colony survival of MRC5 WT cells and BRM knockout clones c6 and c7. Colony number was normalized to untreated conditions. Mean & S.E.M. of four (UV-survival) and two (cisplatin-survival) independent experiments, each performed in triplicate, are presented. *P < 0.05, **P < 0.01, ***P < 0.001, relative to WT, n.s., non-significant. Full-size immunoblot scans are provided in Supplementary Fig. 7c, d