Mitochondrial Oxidative Stress due to Complex I Dysfunction Promotes Fibroblast Activation and Melanoma Cell Invasiveness
Figure 1
Oxygen superoxide level and α-SMA in fibroblasts carrying mitochondrial dysfunctions of complex I. (a) Flowcytometer analysis of level. Control fibroblasts (HFY) and fibroblasts carrying mutations in NDUFS1 gene (Q522K and R557X/T595A), in NDUFS4 gene (W15X), and in the nuclear PINK gene (W437X) were cultured for 48 hours in low glucose medium and then incubated with 5 mM Mitosox for 10 minutes at 37°C for detection of oxygen superoxide. A flowcytometer analysis is then performed. The results are representative of five experiments with similar results. * mutated fibroblasts versus control fibroblasts. (b) Fibroblasts seeded on glass coverslips are treated as in (a) and a confocal microscopy analysis is performed. (c) Analysis of α-SMA expression in control fibroblasts (HFY) and fibroblasts carrying mutations. Lysates of cells were subjected to α-SMA immunoblot analysis. An antiactin immunoblot was performed for normalization. (d) Analysis of α-SMA expression in control fibroblasts (HFY) and fibroblasts carrying mutations seeded on glass coverslips by confocal microscopy analysis.
