Fig. 7

Combined inhibition of MAPK/ERK pathway in HDF. HDF were stimulated with 2 µM of vemurafenib (PLX4032, or PLX for short), 5 nM of trametinib (TR), or the combination. A Flow cytometric analysis of the proliferative state after 18 h of inhibition. Top: quantification of the percentage of proliferative cells (Ki67 positive) for four independent experiments (n = 4). Bottom: representative histograms showing the frequency distribution of Ki67 levels for the indicated condition. B Flow cytometric analysis of cellular viability after 66 h of inhibition. Top: quantification of the percentage of viable cells (Annexin V–PI negative) for four independent experiments (n = 4). Bottom: Representative scatter plots showing the viable (lower left quadrant) and apoptotic/necrotic cells. C MEK1/2 and ERK1/2 phosphorylation after 18 h of inhibition. Right: representative Western blot for the phosphorylated (p-) and total proteins. GAPDH is shown as loading control. Left: densitometry quantification for three independent experiments (n = 3). Optical density (OD) was normalized to the corresponding loading control. All bar plots show the mean ± SEM of the experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001