Fig. 7
Activated P38MAPK induced phosphorylation of ADAM17 in injured cardiomyocyte. A–C Representative western blot (A) and quantitative results depicting the phosphorylation vs. total protein levels of P38 (B) and ERK1/2 (C) in H9C2 cardiomyocytes treated with 600 μM hydrogen peroxide for 12 h (n = 3). D Phosphorylated ADAM17 (p-ADAM17) was assessed by western blot with anti-phospho-ADAM17 (pThr735) antibody after transfection with ADAM17 siRNA or P38MAPK inhibitor SB203580 as indicated (n = 3). E Flow cytometric analysis of annexin V/PI double stained cardiomyocyte after treatment as indicated (right). Histogram indicating the percentage of later apoptosis cells in 4 groups (left) (n = 3). F The cell viability was detected by CCK8 assay (n = 4). G The live (green) and dead (red) cells were observed by calcein-AM/PI double staining kit after treatment as indicated (right). Histogram showing percentage of viable cells(left) (n = 3). H Protein levels of ACE2 (110 kDa), collagen I, collagen III TGF-β1 in cardiomyocytes and released ACE2 protein (90 kDa) in the supernatant were detected by western blotting(n = 3). I, J ACE2 activity in cardiomyocyte lysates (I) and cardiomyocyte supernatant (J) was detected by the ACE2 Activity Fluorometric Assay(n = 4). Data shown as mean ± SEM. Derived by two-sample t-test, *P < 0.05; **P < 0.01, ***P < 0.001