Fig. 3.

Effects of HGF on expression of RAD51 in colon cancer cells. a, b HGF (25 ng/ml) was added to HT-29 and HCT-116 cells for the indicated times. The expression of RAD51 was performed by Western blotting. c HT-29 and HCT-116 cells were treated with 25 ng/ml HGF for 8 h, then subjected to subcellular fractionation. The expression of PARP, Tubulin and RAD51 in the cytosol (C) and nuclear (N) fractions was analyzed by Western blotting. d HT-29 cells were treated with 25 ng/ml HGF for 8 h, and cyto-staining for RAD51 (green) and nuclei (blue) were detected by immunofluorescence. e HCT-116 cells were transiently transfected with scramble control siRNA or two pairs of RAD51 siRNA separately for 48 h. The expression of RAD51, ATR, Chk1, TopBP1, and Chk1 phosphorylation was performed by Western blotting. f HCT-116 cells were transiently transfected with scramble control siRNA or RAD51 siRNA, then added with 1 μM HU for 1 h. The expression of RAD51, Chk1 and Chk1 phosphorylation was performed by Western blotting. g HCT-116 cells were treated with 25 ng/ml HGF for 4 and 8 h. The mRNA levels of RAD51, TopBP1, and GAPDH were measured by RT-PCR. (Color figure online)