Fig. 4.

Effects of AKT on Chk1 phosphorylation. a, b HGF (25 ng/ml) was added to HT-29 and HCT-116 cells for the indicated times. The expression and phosphorylation of AKT was performed by Western blotting. c HT-29 and HCT-116 cells were transiently transfected with Scramble Control siRNA or AKT siRNA separately for 48 h. The expression of AKT, Chk1, ATR and Chk1 phosphorylation was performed by Western blotting. d HT-29 and HCT-116 cells were treated with 25 μM LY294002 for 6 and 16 h. The expression and phosphorylation of AKT and Chk1 was performed by Western blotting. e HT-29 and HCT-116 cells were treated with 25 μM LY294002 for 16 h, then subjected to subcellular fractionation. The expression of PARP, Tubulin and Chk1 phosphorylation in the cytosol (C) and nuclear (N) fractions was analyzed by Western blotting. f HCT-116 cells were transiently transfected with Scramble Control siRNA or AKT siRNA separately for 48 h, then added with 25 ng/ml HGF for 4 h. The expression of AKT, Chk1 and Chk1 phosphorylation of was performed by Western blotting. g HT-29 and HCT-116 cells were pretreated with 25 μM LY294002 for 16 h, then added with 25 ng/ml HGF for 4 h. The expression and phosphorylation of AKT and Chk1 was performed by Western blotting. h HT-29 and HCT-116 cells were pretreated with 25 μM LY294002 for 16 h, then added with 1 μM HU for 1 h. The expression and phosphorylation of AKT and Chk1 was performed by Western Blotting. i HCT-116 cells were transiently transfected with scramble control siRNA or AKT siRNA separately for 48 h, then added with 1 μM HU for 1 h. The expression of AKT, Chk1 and Chk1 phosphorylation was performed by Western blotting