Fig. 2.

POU2F1 promoted CF differentiation and enhanced extracellular matrix protein synthesis. (A−I) CFs were infected with Ad-Ctrl or Ad-POU2F1 then treated with saline or Ang II for 48 h. (A) the expressions of POU2F1, fibronectin, and α-SMA were determined using the Western blot; (B−D) quantitative analysis of the relative protein expressions of (B) POU2F1, (C) fibronectin, and (D) α-SMA; (E) representative images and quantifications of the immunofluorescence analysis of (F) fibronectin, (G) Collagen I, and (H) α-SMA in CFs; (I) the secretion of Collagen I in the cell culture supernatant of CFs was determined by ELISA; (J−R) CFs were infected with Ad-NC or Ad-sh-POU2F1 to knock down POU2F1 then treated with saline or Ang II for 48 h; (J) Western blot analysis of POU2F1, fibronectin, and α-SMA in CFs. Quantitative analysis of relative protein expression of (K) POU2F1, (L) fibronectin, and (M) α-SMA in CFs; (N) Representative images and quantifications of the immunofluorescence analysis of (O) fibronectin, (P) Collagen I, and (Q) α-SMA in CFs; (R) the secretion of Collagen I in the cell culture supernatant of CFs was determined by an ELISA. Data are presented as mean ± SEM. n = 6. A two-way analysis of variance followed by Tukey's post hoc test was used. * p < 0.05; ** p < 0.01; *** p < 0.001. α-SMA = α-smooth muscle actin; Ad-Ctrl = control adenovirus; Ad-NC = negative control adenovirus; Ad-POU2F1 = POU2F1-overexpressed adenovirus; Ad-sh-POU2F1 = sh-POU2F1 adenovirus; Ang II = angiotensin II; CF = cardiac fibroblast; ELISA = enzyme-linked immunosorbent assay; GAPDH = glyceraldehyde-3-phosphate dehydrogenase; POU2F1 = POU domain, class 2, transcription factor 1; SEM = standard error of mean.