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. 2010 Nov 17;25(1):138–156. doi: 10.1210/me.2010-0217

Figure 4.

Figure 4

MC1R-mediated ERK activation in HBL cells is independent of PKC, calcium fluxes, or PKA. A, HBL cells were preincubated for 1 h in the presence or absence of the protein kinase inhibitors PD98059 (50 μm), Ro31–8425 (25 nm), or Rp-cAMP (20 μm) and then challenged with either NDP-MSH (10−7 m, 5 min) or PMA (0.1 ng/μl, 15 min). Cell lysates were analyzed by Western blot for detection of phosphorylation of a PKC substrate (MARCKS, upper blot) and total ERK2 as loading control. B, HBL cells were challenged with NDP-MSH (10−7 m, 5 min) with or without preincubation (1 h) with Ro31–8425 or Rp-cAMP. Cell extracts were blotted for pERK. C, HBL cells were challenged with NDP-MSH (10−7 m, 5 min) with or without (±)-Bay K8644 (1 μm, 1 h) or nifedipine (10 μg/ml, 1 h). ERK phosphorylation was analyzed by Western blot.