Fig. 2.

Effect of FSK (10 μmol/l), in the absence (top bars) and presence (bottom bars) of the PKA inhibitor H89 (2 μmol/l) on JNK phosphorylation in human cultured cutaneous VSMCs. Cells were harvested 5, 10, 20, and 60 min after addition of FSK, and cell lysates were assessed for total and phosphorylated JNK using Western blot analysis. Top: representative blot demonstrating effect of FSK (10 μmol/l), in the presence and absence of H89 (2 μmol/l), or CMC (100 μmol/l) on levels of phospho-JNK or total JNK 10 min after administration of the agonists. When present, H89 was administered 30 min before and during exposure of the cells to FSK. FSK or CMC increased phosphorylation of JNK that was sustained for at least 20 min but had returned to basal levels by 60 min. H89 did not influence the response to FSK at any time point. The results are expressed as fold increase from time 0 and are presented as means ± SE for n = 4 different cell culture experiments. *Significantly different from control; *P < 0.05; **P < 0.01; ***P < 0.001.