Figure 2.

Knockdown of Cfast inhibits myofibroblast differentiation in isolated cardiac fibroblasts

(A) Expression of Cfast after lentivirus-based knockdown in cardiac fibroblasts. Data represent mean ± SEM (n = 6); p values were determined by Student’s t test. (B) Reduced expression of fibrosis genes after Cfast knockdown (Kd) in cardiac fibroblasts. Data represent mean ± SEM (n = 6), p values were determined by Student’s t test. (C) Western blot showing the expression of indicated protein in control and Cfast kd fibroblasts. (D) Quantification of western blot band density. n = 3. p values were determined by Student’s t test. (E) Immunostaining of α-smooth muscle actin (α-SMA) in control and Cfast kd fibroblasts. α-SMA labels activating cardiac fibroblasts (green) and DAPI labels nuclei (blue). (F) Quantification of α-SMA positive cells in control and Cfast kd fibroblasts. p values were determined by Student’s t test. (G) qRT-PCR of the expression of α-Sma, Vimentin, and Periostin genes in control and Cfast kd fibroblasts. p values were determined by Student’s t test. (H and I) Mouse cardiac fibroblasts were transfected with lentivirus-Cfast to overexpress Cfast (Cfast Over) or with lentivirus-GFP (control) for 48 h. Gene expression of Cfast (H) and fibrosis related genes (I), Col1a1, Col3a1, Fn1, Eln, and α-SMA were measured by qRT-PCR. Data represent mean ± SEM (n = 6). p values were determined by Student’s t test.