Fig. 6

Variations in cardiomyocytes in each group treated with 5 μM PRGL493 and 20 μM ammidin. (A-B) Protein expression of ACSL4, AMPK, p-AMPK, mTOR and p-mTOR. Proteins were then transferred onto PVDF membranes. After cutting according to the molecular size, the primary antibody was incubated. (C-D) JC-1 staining showed that JC-1 aggregated in the mitochondria of normal cells emitted red fluorescence, while JC-1 in dead cells emitted green fluorescence due to its presence in the cytoplasm as a monomer. Merge: A combination of aggregate (red fluorescence), monomer (green fluorescence) and nucleus (blue fluorescence). (E) Cardiomyocyte viability. (F) SOD level. (G-H) ROS fluorescence image, Merge: A combination of ROS (green fluorescence) and nucleus (blue fluorescence). (I) LDH release rate. (J) MDA level. (^*P < 0.05 vs. control. ^#P < 0.05 vs. H/R; scale bars: 100 μm; n = 3). ACSL4, Acyl-CoA synthetase long-chain family member 4; p-, phosphorylated; SOD, superoxide dismutase; ROS, reactive oxygen species; LDH, lactate dehydrogenase; MDA, malondialdehyde; H/R, hypoxia/reoxygenation;JC-1,1,1′,3,3′-tetraethyl-5,5′,6,6′-tetrachloroimidacarbocyanine iodide