Fisetin enhanced the viability of H9c2 cardiomyocytes. (A) Survival tests were performed in normoxia, HS or HS/R experiments. Different concentrations of fisetin (from 0 to 100 μM) were tested. In normoxia experiments cells were cultured during 24 h in normal conditions, while in HS experiments cells were cultured in serum free DMEM at 0.5% O2, both in presence of the drug. In the HS/R experiments, cells were treated with fisetin following HS, during the whole reoxygenation phase. Cell survival was monitored by CyQuant assay. Results are expressed as the mean of three independent experiments performed in triplicate (error bars: SEM). (B) Effect of fisetin on cardiomyocytes viability upon HS/R treatment. Cell viability was measured in FACS by annexin V/PI staining: annexin V−/PI− cells were considered as viable. Normoxia: cells cultured under normoxia without fisetin (control group). HS/R: cells subjected to HS/R, treated with DMSO as vehicle control. HS/R + F: cells subjected to HS/R, treated with 15 μM fisetin. Results are expressed as the mean of three independent experiments. One-way ANOVA. Post-hoc analysis by Tukey. *P ≤ 0.05, ****P ≤ 0.0001.
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