Accelerated Recovery of Mitochondrial Membrane Potential by GSK-3β Inactivation Affords Cardiomyocytes Protection from Oxidant-Induced Necrosis
Figure 6
Effects of inhibition of GSK-3β on antimycin A-induced changes in ΔΨm and its recovery.
A: Western blotting for Ser9-phospho- and total GSK-3β in mitochondria, B: Effects of antimycin A (AA) on phospho-GSK-3β level. C: TMRE fluorescence after AA treatment in vehicle- and LiCl-pretreated cells. Western blotting for Ser9-phospho-GSK-3β, total GSK-3β, Ser641/645-phospho-glycogen synthase (GS), non-phospho-GS and β-actin (loading control) in total lysates of vehicle-treated and LiCl-treated cells. Treatments with 30 mM and 60 mM LiCl for 60 min induced phosphorylation of GSK-3β and dephosphorylation of GS. Increased phosphorylation of GSK-3β by LiCl reflects reduced activity of protein phosphatase 1, which is positively regulated by GSK-3β activity. D: TMRE fluorescence after AA treatment in control siRNA- and GSK-3β-siRNA-pretreated cells. NC = nicorandil. Treatment = time after onset of treatment with AA, Washout = time after washout of AA. *p<0.05 vs. Vehicle or Control siRNA. N = 8.