Fig. 6: Mst1, FoxO1, and C/EBP-β play protective roles in response to prolonged ischemia in the heart.
a Proteins from cultured cardiomyocytes exposed to 4 h of hypoxia were detected with an antibody against either C/EBP-β phosphorylated at Thr299 or Mst1 phosphorylated at Thr183. Left: representative images of immunoblot analyses. Right: the results of quantitative analyses (n = 3). b Cardiomyocytes treated with Ad-shMst1 or Ad-shScramble were stained with a C/EBP-β antibody (green), a troponin T antibody (red), and DAPI (blue). c Cardiomyocytes were transduced with 10 MOI of adenovirus harboring either shMst1 or shScramble in indicated combinations. Seventy-two hours after the transduction of adenoviruses, some samples were exposed to hypoxia for 4 h, then cardiomyocytes were harvested. The survival of cardiomyocytes was quantitated by CellTiter-Blue assay. The experimental data were normalized by values obtained from control cardiomyocytes without adenovirus transduction (n = 12). d Proteins in fractionated cellular lysates from cultured cardiomyocytes exposed to indicated periods of hypoxia were detected with Mst1, GAPDH (cytosolic marker), and Histone H3 (nuclear marker) antibodies. e Tg-Mst1 (n = 6), Tg-DN-Mst1 (n = 5), C/EBP-β+/− (n = 6), C/EBP-β+/− crossed with Tg-Mst1 (bigenic) (n = 6), FoxO1-cKO (n = 5), and WT (n = 8) mice were subjected to prolonged ischemia for 4 h. Upper: gross appearance of LV myocardial sections after Alcian blue and TTC staining. Lower left graph: comparison of AAR (percentage of LV) among indicated groups. Lower right graph: comparison of infarct size/AAR among indicated groups. 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) or one-way ANOVA followed by Tukey’s multiple comparison test in (b, e). Data are mean ± SEM. Source data are provided as a Source Data file.
