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. 2017 Aug 22;11:253. doi: 10.3389/fncel.2017.00253

Figure 4.

Figure 4

Downstream activation of extracellular-signal-regulated kinase (ERK)1/2 does not mediate the effect of PKC activation on NET activity. Naïve or MKP3-expressing (PC12-MKP3) PC12 cells were pretreated for 30 min with PKC activator PMA (1 μM), MEK1/2 inhibitor PD98056 (20 μM), or EGF (100 nM) and NET-specific radiolabeled uptake was performed subtracting desipramine insensitive background uptake (A). All experiments were performed in triplicate in three independent experiments. Statistical significance was determined employing one-way ANOVA with Bonferroni post hoc analysis (***p < 0.001 comparing effects of PMA treatment. There was no significant effect of PD98056 on the effect of PMA treatment and no significant effect of EGF treatment compared to vehicle). Identical pharmacologic treatments of the same cell types were performed and ERK1/2 map kinase activation was assayed by immunoblotting (B). PMA showed small activation of ERK1/2 and this was inhibited by PD98056. EGF showed strong activation of ERK1/2.