Figure 7.

TGFβ1 via Prrx2‐Wnt5a signalling induces cell differentiation in cardiac fibroblasts. A‐D, Cultured cardiac fibroblasts were co‐infected with adenovirus expressing Prrx2 shRNA plus vector or Wnt5a cDNA for 48 h and then treated with TGF‐β (10 ng/mL) for 24 h. Cell differentiation was determined by immunofluorescence analysis of α‐SMA and quantitative analysis was shown in A. Gene expressions of Wnt5a, α‐SMA, GAPDH, collagen I (Col I) and collagen III (Col III) in cells were measured by real‐time PCR in B. Total cell lysates of cardiac fibroblasts were subjected to perform Western blotting analysis to detect protein levels of Wnt5a, α‐SMA, Col I, and Col III in C and quantitative analysis was performed in D. N is 5 in each group. *P < .05 vs Vector. E‐H, Cardiac fibroblasts were co‐infected with adenovirus expressing Prrx2 cDNA plus negative control (NC) shRNA or Wnt5a shRNA for 48 h. Cells were subjected to detect immunofluorescence analysis of α‐SMA and quantitative analysis was shown in E, gene expressions of Wnt5a, α‐SMA, GAPDH, Col I and Col III in F, protein levels of Wnt5a, α‐SMA, Col I, and Col III in G and H. N is 5 in each group. *P < .05 vs NC plus Prrx2