Figure 10

Both PAR1 and PAR2 are involved in the hypertrophic effect of KLK8.

Primary cultured neonatal cardiomyocytes were infected with KLK8 adenovirus, 24 h later control siRNA, PAR1 siRNA or PAR2 siRNA was transfected into the cardiomyocytes. Twenty four hours later, all the media of cells were replaced with fresh media, then PAR1 antagonist RWJ56110, or PAR2 antagonist FSLLRY-NH2 was added into the culture media. After incubation for 24 h, cells were treated with PE for another 48 h. (A) showed KLK8 protein expression. (B) total protein content was determined by BCA assay; (C) Cardiomyocytes were stained with FITC-conjugated phalloidin as described in “Methods”. One hundred cells from randomly selected fields in three independent expreriments were evaluated for cell size using the Image-Pro Plus cell area measurement software. (D–F) transcripts of cardiac hypertrophy markers including ANP (D), BNP (E) and Myh7 (F) were determined by quantitative real-time RT-PCR. Values are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01 vs cells treated with Ad-vector and control siRNA; ^#P < 0.05, ^##P < 0.01vs cells treated with Ad-KLK8 and control siRNA.