FIGURE 1.

Brg1 is required for efficient removal of CPD but not 6-4PP. A, knockdown of Brg1 level by hBrg1 siRNA. OSU-2 cells were transfected with Brg1 siRNA (20 nm) using Lipofectamine 2000. Cell lysate was collected 24 or 48 h after transfection and subjected to Western blot analysis with anti-Brg1 antibody. Lamin B was detected to serve as an internal control. B and C, effect of Brg1 on the efficiency of CPD (B) or 6-4PP (C) removal. After 24 h of transfection with Brg1 siRNA, OSU-2 cells were cultured in serum-free medium for an additional 24 h. Cells were UV irradiated at 10 J/m² and allowed to repair for indicated times. The same amount of genomic DNA was subjected to ISB analysis and the amount of CPD (B) or 6-4PP (C) was detected with anti-CPD or anti-6-4PP antibody, respectively, at each time point. Data presented in the graph on the right represent the relative CPD or 6-4PP remaining in each sample. Percentage CPD or 6-4PP was calculated from the intensity relative to initial irradiated sample. The data points represent an average of three independent measurements, with error bars representing standard deviation.