Fig. 2

Characterization of NSC12^res/NSC12^sens UM subpopulations. A Semi-quantitative PCR analysis of FGF and FGFR expression in control and NSC12^res/NSC12^sens subpopulations of 92.1 (left panels) and Mel270 (right panels) cells. B Western blot analysis of phospho-FGFRs and FGFR1 in control and NSC12^res/NSC12^sens subpopulations of 92.1 (left panels) and Mel270 (right panels) cells. The lower panels show the densitometric analysis of immunoreactive bands normalized to GAPDH protein levels. Data are the mean ± SEM of three independent experiments. *p < 0.05 and **p < 0.01 vs control; ^#p < 0.05 vs NSC12^res, ANOVA. C Human Phospho-Kinase Antibody Array on 92.1_NSC12^res and 92.1_NSC12^sens total cell lysates. The heatmap shows the color-coded normalized protein levels of all detected phospho-proteins. The lower panels show the spots of p38∝, ERK1/2, JNK1/2/3, GSK-3∝/β, CREB, HSP27, RSK 1/2/3, p70 S6 Kinase and FAK and the corresponding densitometric analysis (relative units, RU). Data are the mean ± SEM of two technical replicates in one representative experiment out of three independent measurements that provided similar results. **p < 0.01 and ***p < 0.001 vs NSC12^res, Student’s t-test. D The production of mtROS was assessed on control and NSC12^res/NSC12^sens subpopulations of 92.1 (left panels) and Mel270 (right panels) cells using the fluorescent probe MitoSox by cytofluorimetric analysis. In the plots the black line refers to the gate (P1/P2) setting the positive/negative cell populations. Data are the mean ± SEM of three independent experiments. *p < 0.05 and **p < 0.01 vs control; ^#p < 0.05 vs NSC12^res, ANOVA