FIGURE 4:

ROCK1 regulates Rnd3 interaction with 14-3-3 during apoptosis. (A) DLD1 cells coexpressing FLAG-14-3-3β and GFP-Rnd3 were treated with 250 ng/ml anti-Fas antibody and 10 mg/ml cycloheximide for 3 h to induce apoptosis in conjunction with either 10 µM Y-27632 or 50 µM Z-VAD-FMK. Lysates were prepared and immunoprecipitated with anti-FLAG antibody-conjugated beads and the precipitates were probed with anti-GFP or anti-FLAG antibodies. GFP-Rnd3 (co-IP) and GFP-Rnd3 (input) signals were quantified by densitometry to obtain the ratio co-IP to input. The signal ratio of control cells was set to 1. Results shown are representative of five independent experiments. (B) The means of N = 5 independent experiments represented by A are shown with the SD. *P < 0.05, ***P < 0.001 (one-way ANOVA). (C) A model describing the regulation of Rnd3 activation by ROCK1 and 14-3-3 during apoptosis.