Fig. 3

SIRT3 overexpression attenuates inflammation in human cardiac cells. a Relative quantification of IL6, MCP1, and SOCS3 mRNA expression in human AC16 cardiac cells transfected with LacZ-carrying or SIRT3-carrying plasmids in the presence or absence of TNF-α (TNF, 10 ng/mL, 24 h). The graphs represent the quantification of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-normalized mRNA levels, expressed as a percentage of the control samples ± SD. b Western blot analysis showing the levels of IκBα in total protein extracts, p65 in nuclear protein fractions, and FOS in cytosolic (CP) and nuclear (NE) protein fractions obtained from the same samples. The graphs represent the quantification of protein levels normalized to actin (total protein) or lamin B (nuclear protein), expressed as a percentage of the control samples ± SD. c EMSA data showing NF-κB DNA-binding activity. Ab antibody; IC immunocomplex; NE nuclear extract. d EMSA data showing AP-1 DNA-binding activity in the same samples depicted in panel a. e Relative quantification of ATF4, EDN-1, and TGFB1 mRNA expression expressed as a percentage of the control samples ± SD. The data were compared by ANOVA followed by Tukey’s post hoc test. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. LacZ; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. LacZ + TNF-α. f Pearson correlation coefficient between SIRT3 and FOS gene expression in left ventricular myocardial tissue obtained from patients undergoing aortic valve replacement surgery. The relative transcript levels of the target genes, in arbitrary units, were used to calculate the Pearson correlation coefficient