Fig. 7: C/EBP-β-T250E knock-in mice are protected against myocardial ischemia.

a C/EBP-β-T250E knock-in (C/EBP-β-KI) and WT mice were subjected to 45 min of ischemia and 24 h of reperfusion. Upper: the gross appearance of LV myocardial sections after Alcian blue and TTC staining. Lower left graph: comparison of the size of the AAR (percentage of LV) in the two groups (n = 5). Lower right graph: comparison of the infarct size/AAR in the two groups (n = 5). b Quantitative analyses of left ventricular ejection fraction in C/EBP-β-KI and WT mice after 45 min of ischemia and 24 h of reperfusion (n = 3). c qRT-PCR analyses of RNA extracted from heart homogenates of C/EBP-β-KI or WT mice to evaluate the effect of C/EBP-β phosphorylation at Thr250 upon mRNA expression of cell death- and survival-associated genes, including PRDX2 (n = 8), TRAIL (n = 5), catalase (n = 7), NOXA (n = 7), FasL (n = 3), MnSOD (n = 5), and Bnip3 (n = 3). d Chromatin immunoprecipitation assays with sequencing to examine the effect of C/EBP-β phosphorylation at Thr299 upon the binding ability of C/EBP-β and FoxO1 to the promoter regions of Cat and Mnsod genes during myocardial ischemia. Upper two panels indicate the binding ability of CEBP-β to the promoter regions of the Cat and Mnsod genes in WT and C/EBP-β-KI ischemic hearts. Lower two panels indicate the binding ability of FoxO1 to the promoter regions of the Cat and Mnsod genes in WT and C/EBP-β-KI ischemic hearts. e Motif enrichment analyses showed similarities in binding sequences of C/EBP-β and FoxO1. All experiments were repeated at least three times, with n representing biologically independent replicates. P values were determined by two-sided unpaired Student’s t test in (a, c) or one-way ANOVA followed by Tukey’s multiple comparison test in (b). Data are mean ± SEM. Source data are provided as a Source Data file.