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. 2013 Jun 11;8(6):e66029. doi: 10.1371/journal.pone.0066029

Figure 8. Phosphorylation at Ser96 in RhoGDI1 is required for CCK-induced RhoA and Rac1 activation.

Figure 8

WT-RhoGDI1 and the phosmomimetic mutants S34D-RhoGDI and S96D-RhoGDI were expressed in pancreatic acini using adenoviral delivery. The expression of GFP was used as a control. Overnight incubated acini were stimulated with CCK (300 pM) for 5 min and then RhoA activation or 10 min and then Rac1 activation were determined using a pull-down assay. Top: A representative immunoblot shows that the expression of WT-RhoGDI1 inhibits CCK-induced RhoA activation or Rac1 activation. Only the expression of phosphomimetic mutant S96D-RhoGDI1 induced CCK-stimulated GTP-RhoA and GTP-Rac1 levels (Representative of 4 experiments). Comparable expression of WT-RhoGDI1, as well as mutants was analysed by Western-blotting using an anti-RhoGDI1 antibody. Bottom: Quatitative analysis of either RhoA or Rac1 activation. Values are means ± SE (n  = 4 experiments). *: p<0.05 vs control and †: p<0.05 vs CCK.