VASP Activation via the Gα13/RhoA/PKA Pathway Mediates Cucurbitacin-B-Induced Actin Aggregation and Cofilin-Actin Rod Formation
Figure 3
H89, but not MDL12330A (MDL), pretreatment suppressed CuB-induced VASP activation, actin aggregation and cell deformation.
(A) Western blotting showing the effects of H89 and MDL pretreatment on CuB-induced VASP activation and cofilin dephosphorylation. (B,C) CuB decreased the levels of cAMP in a time- and dose-dependent manner (B). H89 and MDL pretreatment affected the levels of cAMP in CuB-treated cells (C). The cAMP level of forskolin (10 μM)-treated cells was set as 100% activity (B,C). (D) Immunofluorescence analysis of the influence of H89 pretreatment on the distribution of VASP (red) and actin (green) in CuB-treated cells. Scale bars: 10 μm (5 μm in magnified images). (E) Co-localization analysis of VASP and actin of D. (F) Effect of H89 pretreatment and MDL pretreatment on CuB-induced cell deformation as observed using phase-contrast microscopy. Magnified images of the boxed areas are presented on the right. Scale bars: 20 μm.