TNF-α receptors knockout animals showed decreased basal antioxidant gene/proteins in the heart. (A) One step RT-PCR analysis of TNF-α receptors (TNFR1 and TNFR2) and housekeeping gene Gapdh were determined in WT and TNFR DKO heart samples. (B) Quantitative real time PCR analysis of TNFα receptors (TNFR1 and TNFR2) and housekeeping gene Gapdh were determined in WT and TNFR DKO heart samples (n=4). (C) Nrf2 and Keap1 gene expression profiles were measured by qPCR. Relative gene expression was calculated by normalizing the mRNA levels of gene of interest with Gapdh. (D) Nrf2 regulated antioxidant proteins (NQO1, HO-1, G6PD, SOD1, SOD2 and CAT) were determined by Immunoblot analysis in WT and DKO heart tissue homogenates (n=4). (E) Densitometric analysis of proteins normalized with GAPDH intensity using Image J. Statistical differences were determined by Mann Whitney test, where *p<0.05 vs control.