Fig. 7.

PHB2 overexpression alleviated DOX-induced mitochondrial dysfunction through NDUFV2 in cardiomyocytes.

(A) Representative immunoblotting images of PHB2 overexpression and NDUFV2 knockdown using adenoviruses in primary cardiomyocytes (β-Tubulin used as an internal control). (B and C) OCR curves and quantification of basal respiration, maximal respiration, and ATP-coupled respiration in primary cardiomyocytes with and without DOX challenge (0.1 μM) for 24 h after adenovirus transfection (n = 6 independent experiments per group). (D) ATP production rate in primary cardiomyocytes under various treatment settings (n = 6 independent experiments per group). (E and F) Representative images and quantification of fluorescence intensity of MitoSOX staining in AMCMs under various treatment settings (n = 20 independent experiments per group). (G and H) Representative images and quantification of fluorescence intensity of TMRM staining in AMCMs under various treatment settings (n = 20–30 independent experiments per group). (I) Measurement of cell viability using CCK-8 and CellTiter-Glo luminescent assay in primary cardiomyocytes under various treatment settings (n = 6 independent experiments per group). (J) LDH leakage in supernatants of primary cardiomyocytes under various treatment settings (n = 6 independent experiments per group). (K, L, and M) Representative immunoblotting images and quantification of FIS1, OPA1, and MFN2 in primary cardiomyocytes under various treatment settings (β-Tubulin used as an internal control, n = 4 independent experiments per group). Mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, no significance. For statistical analysis, two-way ANOVA with Tukey's test for multiple comparisons was used.