Figure 6.

Effects of cyclic stretch on PUMA binding and promoter activity in cardiomyocytes. (A) Representative EMSA showing protein binding to IRF-1 oligonucleotide in nuclear extracts of cardiomyocytes following cyclic stretch during various times and in the absence or presence of JNK inhibitors or siRNA, IFN-γ antibody or addition of 200 pg/ml IFN-γ. Arrows indicate the mobility of the complex. Similar results were found in another two independent experiments. Cold oligo means unlabeled IRF-1 oligonucleotide. (B) Constructs of PUMA promoter gene. (C) Quantitative analysis of PUMA promoter activity. Cardiomyocytes were transiently transfected with pPUMA-Luc by a gene gun. The luciferase activity in cell lysates was measured and was normalized with Renilla activity (n = 3 per group). *P < 0.01 vs. control.