Figure 5. GnRH-induced ERK phosphorylation and nuclear localization is partially dependent on PKC activation.

Cells were treated, imaged and analysed as described for fig. 2, except that the stimulation was for 10 min with 1 µM PDBu or 10 nM EGF in cells that had been pre-treated for 10 min with DMSO vehicle (Ctrl), 200 nM Ro31-8425 PKC inhibitor, 100 nM AG1478 EGFR inhibitor or 10 µM PD184352 MEK inhibitor (panel A), or the cells were stimulated for 5 min with the indicated concentrations of GnRH in the presence of the same inhibitors (panel B). Panel A show population average values for ppERK in AFU (top panel) and ERK N:C (bottom panel) after treatment with inhibitors and agonists as indicated. Data are shown are mean ± SEM (n = 3), ** = p<0.01, comparing Ctrl and inhibitor co-incubations for each readout, according to one-way ANOVA and Dunnet’s post-hoc test. Panel B shows concentration-response curves for the effect of GnRH on population averaged ppERK intensity (in AFU) and ERK N:C responses. Data are shown means ± SEM (n = 6) and two way ANOVAs revealed GnRH, Ro31-8425 and PD184352 as significant sources of variation (P<0.01) for both measures (ppERK and ERK N:C), whereas AG1478 was not a significant variable for either (P>0.05). *P<0.05, **P<0.01 comparing with and without inhibitor at matched GnRH concentration.