Figure 5.

Effect of AMPK inhibition on melatonin-mediated regulation of apoptosis and mitochondrial membrane potential. Cells in each group were treated for 12 h with 4 mmol/L Ox, 10 µmol/L MLT, and/or 5 µmol/L Cpd C 2HCl. (a) Flow cytometry was used to detect HK-2 cell apoptosis. (b) Immunofluorescence of Mito-Tracker Red CMXRos-staining (red) (200× magnification) to determine HK-2 cell mitochondrial membrane potential of the respective groups, (c) Immunofluorescence of JC-1 (200× magnification) to determine HK-2 cell mitochondrial membrane potential in the respective groups, (d) ELISA showing caspase-3 activity. Stronger red fluorescence signal correlates with higher mitochondrial membrane potential. Data are presented as the mean ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 versus the NC group; #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001 versus the Ox group; ^P < 0.05, ^^P < 0.01, ^^^P < 0.001, ^^^^P < 0.0001 versus the Ox + MLT group