Effect of hnRNP C depletion on cell cycle distribution before and after IR.

<p><b>A</b>. DR-U2OS cells were treated with control, PALB2 or hnRNP C siRNAs for 72 hr and then subjected to 10 Gy of IR. Cells were labeled with BrdU either before or 16 hr post IR, and cell cycle profiles were analyzed by anti-BrdU staining and FACS. Cells in S, G1 and G2/M phases were indicated by upper, lower left and lower right boxes, respectively. Early S and late S phase cells are separated by an arbitrary dotted line and indicated by “ES” and “LS”. <b>B</b>. Quantification of cell cycle distributions in two independent experiments. Error bars represent standard deviations, and the asterisk indicates p≤0.05. <b>C</b>. Cells were treated with the siRNAs and subjected to IR in the same way as in A, and mitotic index was measured by phosphorylated histone H3 staining and FACS. <b>D.</b> Cells were treated with control or hnRNP C siRNAs for 72 hr and then subjected to 10 Gy of IR. Cells were harvested at indicated time points, and cellular abundance of hnRNP C and γH2A.X were analyzed by Western blotting. <b>E.</b> Cells treated with siRNAs and IR as in <b>D</b> were fixed and the abundance and localization of hnRNP C and γH2A.X were analyzed by IF.