Figure 2.

Transcriptional regulation of fibronectin by DDR2-activated YAP transcription co-activator. (A) Cardiac fibroblasts were treated with verteporfin (10 µM) for 1 h prior to Ang II treatment. Cells were collected at 12 h post-Ang II treatment, and fibronectin protein levels were examined, with β-actin as loading control. ** p < 0.01 (comparisons as depicted in the Figure). (B) Cardiac fibroblasts were transiently transfected with YAP siRNA (10 pmol) or control (scrambled) siRNA prior to treatment with Ang II for 12 h. Fibronectin protein expression was examined, with β-actin as loading control. Validation of DDR2 silencing is also shown. ** p < 0.01 (comparisons as depicted in the Figure). Data are representative of three independent experiments, n = 3, Mean ± SEM. (C) Sub-confluent quiescent cultures of cardiac fibroblasts were transiently transfected with YAP siRNA (10 pmol) or control (scrambled) siRNA prior to treatment with Ang II (1 µM), and another set of cells was pre-treated with WRG-28 for 1 h. Cells were collected at 30 min post-Ang II treatment and chromatin was immunoprecipitated with anti-YAP antibody, followed by PCR amplification, and analyzed on a 2% agarose gel for presence of the 150 bp region of the fibronectin gene promoter containing the TEAD consensus binding site.