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. 2011 Jan;50(1):87–98. doi: 10.1016/j.yjmcc.2010.10.010

Fig. 2.

Fig. 2

Selective activation of PKA and Epac signalling with N6-Benzoyl-cAMP and 8-CPT-2′-O-Me-cAMP, respectively. VSMC were serum deprived for 24 h and stimulated with 200 μM BNZ, CPT, BNZ plus CPT or 100 μM forskolin for 30 min as indicated. A, Cell lysates were analysed for PKA activity by kemptide phosphorylation assay. Upper bands represent PKA phosphorylated kemptide while lower bands are un-phosphorylated kemptide. Recombinant PKA catalytic subunit and lysis buffer were included in the assay as positive and negative controls respectively. B, Cell lysates were analysed for phospho-VASP (slower migrating band) by western blotting with a total VASP antibody, n = 3. C, Rap1 activity was quantified by Ral-GDS pulldown, n = 6. * indicates p < 0.05 versus control.