Figure 2.

Effect of Baf200 depletion on the DNA damage response. A, U2OS cells were treated with the indicated siRNA, followed by treatment with 10 μm etoposide for 20 min to induce damage, and allowed to repair DSBs for the indicated times. Representative images of cells immunostained with DAPI (nuclei) and γH2AX antibodies (DSBs) are shown. The scale bar represents 10 μm in all images. B, quantitative analysis of γH2AX foci formation and resolution after etoposide exposure from three independent experiments is shown. C, quantitative analysis of γH2AX foci formation and resolution after cells were treated with siRNA control and siRNA Baf200 and exposed to ionizing radiation from three independent experiments is shown. D, quantitative analysis of γH2AX foci formation and resolution after cells were treated with different siRNAs targeting Baf200 and exposed to etoposide to induce DNA damage. E, cell cycle distribution of U2OS cells before and after siRNA treatment. U2OS cells were transfected with the indicated siRNAs. After 72 h, cells were stained with propidium iodide, and the percentage of total U2OS cells at G₂, S, and G₁ cell cycle stages was measured by ArrayScan quantification. The data shown are from a single representative experiment out of three repeats; n = 10,000 cells analyzed from a single experiment. The mean ± S.D. is shown.