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. 2014 Sep 3;289(42):29285–29296. doi: 10.1074/jbc.M114.563924

FIGURE 5.

FIGURE 5.

VDAC2-dependent mitochondrial translocation of GSK-3β and mitochondrial production of superoxide. A and B, mRNA levels in H9c2 cells (A) and protein levels in HEK293 cells (B) of VDAC1, VDAC2, and VDAC3 at 48 h after transfection of VDAC2 siRNA. *, p < 0.05 versus control siRNA. C, mitochondrial localization of GFP-tagged GSK-3β (WT) expressed as a percentage of GFP-positive mitochondria among total mitochondria at baseline and at 1 and 3 min after exposure to H2O2 in H9c2 cells. *, p < 0.05 versus baseline; †, p < 0.05 versus control siRNA. D, HEK293 cells were transfected with VDAC1 siRNA, VDAC2 siRNA, or VDAC3 siRNA. Mitochondrial localization of GFP-tagged GSK-3β (WT) expressed as a percentage of GFP-positive mitochondria (mito) among total mitochondria with or without exposure to H2O2 (10 μmol/liter; 3 min) is shown. *, p < 0.05 versus baseline; †, p < 0.05 versus VDAC2 siRNA. E, effects of VDAC2 and kinase activity of GSK-3β on generation of superoxide. Superoxide signal after exposure to H2O2 in H9c2 cells transfected with WT, S9A, or K85R is shown. F, quantitative analysis of superoxide-positive pixels per cell is shown. *, p < 0.05 versus control siRNA; †, p < 0.05 versus WT. Error bars represent S.E. a.u., arbitrary units.