Figure 2: Effects of NGR1 on H/R-induced cell death, depolarization of mitochondrial membrane and intracellular ROS accumulation in H9c2 cardiomyocytes.

H9c2 cardiomyocytes were incubated with indicated concentration of NGR1 (2.5, 5, 10, 20, 40 and 80 μM) for 24 h and then exposed to 6 h of hypoxia and 12 h of reoxygenation. (a) NGR1 had no toxic effect on cell viability with concentrations under 20 μM; Effects of NGR1 on H/R-induced cell viability were detected by (b) MTT and cell death by (c) extracellular LDH; (d) Intracellular ROS levels evaluated using a FACSCalibur flow cytometer; (e) Bar diagram showing intracellular ROS level in H9c2 cardiomyocytes; (f) Bar graphs and (g) representative images of JC-1 red/green cells and merges showed that NGR1 increased the ratio of red to green fluorescence intensity. #P < 0.05 versus the control group, ##P < 0.01 versus the control group, ###P < 0.001 versus the control group, *P < 0.05 versus the H/R group, **P < 0.01 versus the H/R group, ***P < 0.001 versus the H/R group.