Figure 7.

Fingolimod alleviates cardiomyocyte damage induced by H/R. a. Cell viability assessed by CCK-8 assay; n=8. b. Measurement of ROS levels in H9C2 cells using DCFH-DA probe and analysis of mean fluorescence intensity; n=5. c. The MMP was assessed using the JC-1 probe, and the ratio of red to green fluorescence was calculated. Green fluorescence represents JC-1 monomers, while red fluorescence represents JC-1 aggregates; n=5. d. The mitochondrial morphology was examined using TEM, and the Flameng score was calculated based on the observed mitochondrial morphology; n=5. e. Western blot analysis and quantification of S1PR1, S1PR2, and S1PR3 levels in the membranes of H9C2 cells; NaKATPase loading control; n=5. f. The levels of p-STAT3, STAT3, SLC7A11, GPX4, and MnSOD, along with nuclear p-STAT3, were analyzed using Western blot in H9C2 cells, with quantification of results; β-actin and LaminB loading control; n=5. g. The expression of Slc7a11 mRNA in H9C2 cells was quantified using qRT-PCR; n=5. h. The expression of Gpx4 mRNA in H9C2 cells was quantified using qRT-PCR; n=5. i. The expression of MnSOD mRNA in H9C2 cells was quantified using qRT-PCR; n=5. j. Determination of GSH content in H9C2 cells; n=5. k. Determination of Fe²⁺ content in H9C2 cells; n=5. l. Determination of MDA content in H9C2 cells; n=5. All data are means ± standard deviations. Statistical analysis involved one-way ANOVA followed by Tukey's post-hoc test. Lines indicate comparisons between samples, and asterisks denote statistical significance (*P < 0.05, **P < 0.01, ***P < 0.001).