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. 2020 Apr 27;10(12):5623–5640. doi: 10.7150/thno.44836

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PPAR-α knockdown suppresses myocardial FAO levels and protects PP2Cm KO cardiomyocytes against injury following H/R. (A) Expression of PPAR-α by western blotting in cardiomyocytes with or without shPpara adenovirus infection (n=6). (B to N) Ventricular myocytes isolated from WT mice or PP2Cm KO mice were infected with scrambled or shPpara adenovirus for 48 h with or without H/R injury. (B to G) FAO levels were determined by seahorse analyzer (n=4-5). (B) OCR curve treated as mentioned above were determined. (C) Basal respiration (D) ATP production (E) maximal respiration (F) basal respiration due to exogenous FAs and (G) maximal respiration due to exogenous FAs were calculated according to instruction. (H) Expression of ACAA2, ACADM, CD36, CPT1B in cardiomyocytes by western blotting (n=6). (I) Annexin V and propidium iodide (PI) staining by flow cytometry for cardiomyocyte apoptosis determination (n=6). (J) Cleaved and non-cleaved caspase-3 by western blotting (n=6). (K) Cell death assessed by LDH release (n=6). (L) Superoxide production detected by DHE staining (n=6). Scale bar: 20 μm. (M and N) Lipid peroxidation determined by MDA and 4-HNE contents (n=6). Data were analyzed by one-way ANOVA, followed by a Bonferroni post-hoc test. * P<0.05, ** P<0.01. All values are presented as mean ± SEM.

PPAR-α knockdown suppresses myocardial FAO levels and protects PP2Cm KO cardiomyocytes against injury following H/R. (A) Expression of PPAR-α by western blotting in cardiomyocytes with or without shPpara adenovirus infection (n=6). (B to N) Ventricular myocytes isolated from WT mice or PP2Cm KO mice were infected with scrambled or shPpara adenovirus for 48 h with or without H/R injury. (B to G) FAO levels were determined by seahorse analyzer (n=4-5). (B) OCR curve treated as mentioned above were determined. (C) Basal respiration (D) ATP production (E) maximal respiration (F) basal respiration due to exogenous FAs and (G) maximal respiration due to exogenous FAs were calculated according to instruction. (H) Expression of ACAA2, ACADM, CD36, CPT1B in cardiomyocytes by western blotting (n=6). (I) Annexin V and propidium iodide (PI) staining by flow cytometry for cardiomyocyte apoptosis determination (n=6). (J) Cleaved and non-cleaved caspase-3 by western blotting (n=6). (K) Cell death assessed by LDH release (n=6). (L) Superoxide production detected by DHE staining (n=6). Scale bar: 20 μm. (M and N) Lipid peroxidation determined by MDA and 4-HNE contents (n=6). Data were analyzed by one-way ANOVA, followed by a Bonferroni post-hoc test. * P<0.05, ** P<0.01. All values are presented as mean ± SEM.