Fig. 2.

Activation of PKA, but not EPAC1, promotes AP1-associated c-Jun activation in HUVECs. HUVECs were stimulated for 5 h with either 007 (10 μM), the PKA-activator, 6-Bnz (10 μM), or F/R. Nuclear extracts were then prepared and precipitated with the biotinylated oligonucleotides detailed in the upper panel. Cell extracts (input) and precipitated proteins were then Western blotted with the indicated antibodies. Results are representative of an experiment carried out on three separate occasions and densitometic analysis of multiple immunoblots is displayed as a histogram in the lower panel as the ratio of pJun (Ser 63) to total Jun immunoreactivity. Significant differences relative to diluent-treated cells are indicated, ^∗∗∗P < 0.001, as is the significant decrease in immunoreactivity in samples from 6-Bnz-treated cells relative to F/R-treated cells, ^###P < 0.001 (n = 3).