Fig. 2. siRNA-mediated depletion of endogenous RhoG markedly attenuated sustained activation of Rac1 in INS-1 832/13 cells. Panel a: INS-1 832/13 cells were transfected with control-siRNA or RhoG-siRNA. Cell lysates were analyzed by Western blotting for the expression of RhoG. Actin was used as loading control. A representative blot from n=3 independent studies is shown here. Panel b: Pooled data from three independent experiments was shown. Data are expressed as mean ± SEM from three experiments. The data are expressed as fold change relative to Mock. (n=3; * p< 0.05). Panel c: INS-1 832/13 cells were transfected with control-siRNA or RhoG-siRNA. After 48 hours of transfection, cells were subjected to overnight starvation and then were treated with LG (2.5 mM) or HG (20 mM) for 24 hours. Rac1 activation was quantified by Rac1 pull down assay. Representative blots from three independent studies are provided. Panel d: Densitometric quantitation of activated Rac1 in Panel a is shown here. The results from three independent experiments are presented as means ± SEM. The data are expressed as fold change relative to LG-mock (n=3; * p< 0.05). Comparisons shown: a - significant compared with LG-treated mock; b - significant compared with HG treated Mock; c - significant compared with LG treated Cont-si; d - significant compared with HG treated Cont-si.