Figure 6
From: Lats2 promotes heart failure by stimulating p53-mediated apoptosis during pressure overload
Lats2 upregulates Bax and Bak in a p53-dependent manner. (A) and (B), Cultured neonatal rat ventricular cardiomyocytes were transduced with adenovirus harboring either LacZ or Lats2. (A), Protein levels of p53, Lats2 and GAPDH were evaluated with immunoblot analyses. (B), mRNA levels of p53 were evaluated with qPCR analyses. (C), Cultured neonatal rat ventricular cardiomyocytes were transduced with adenovirus harboring either shRNA Lats2 or shRNA control. mRNA levels of p53 were evaluated with qPCR analyses. ***p < 0.001 versus Adenovirus harboring shRNA control. (n = 6). (D), Cultured neonatal rat ventricular cardiomyocytes were transfected with p53-luciferase reporter together with expression plasmids harboring either GFP (control) or Lats2 (left) and either shRNA-control or shRNA-Lats2. Relative luciferase activity is shown. The level of reporter gene activity in control (GFP or shRNA control) transfected cells was designated as 1. (E–G), Cultured neonatal rat ventricular cardiomyocytes were transduced with adenovirus harboring LacZ or Lats2 in the presence of Pifithrin-α (PFT-α), a p53 inhibitor, or dimethylsulfoxide (DMSO), a vehicle. (E), The levels of Bax, Bak, p53, Lats2 and GAPDH in cardiomyocytes were evaluated with immunoblot analyses. Representative images of immunoblot analyses are shown. Experiments were repeated three times. (F), Cardiomyocyte apoptosis was evaluated with TUNEL assays. (G), Cell viability was evaluated with CellTiter-Blue assays. In (B) and (C), *p < 0.05 versus LacZ-transduced cardiomyocytes in the presence of DMSO. #p < 0.05 versus Lats2-transduced cardiomyocytes in the presence of DMSO.
