Figure 4.
PKA and EPAC act synergistically to induce nuclear translocation of CRTC2 and CRTC3. VSMC were transfected with GFP-CRTC 2 (n = 4; A and B) or GFP-CRTC3 (C and D) by electroporation and serum starved for 24 h. Cells were stimulated for 1 h with 2 µM 8-CPT-2′-O-Me-cAMP-AM to activate EPAC, 4 µM 6-Bnz-cAMP-AM to activate PKA or both together. Cells were analysed for cellular localisation of CRTC proteins by fluorescence microscopy and image analysis. Data were expressed as % of transfected cells with nuclear CRTC2 (B) or CRTC3 (n = 3; D) protein. *Indicates p < 0.05, ***indicates p < 0.001 relative to control; one-way repeated measures ANOVA with Student Newman Keuls post-hoc tests.