Fig. 6
Effect of siRNA-mediated knockdown of TFAP4 on differentiation of bovine satellite cells. Cells were transfected with a negative control siRNA (CTRL) or a bovine TFAP4 mRNA-specific siRNA (siTFAP4) and then induced to differentiate into myofibers for 4 days. A Representative images of cells on days 0, 1, 2, and 4 of differentiation. B Representative images of immunostained cells on day 4 of differentiation. Staining of myosin heavy chain protein is shown in green color, and staining of nuclei is shown in blue color. C Fusion index of cells on day 4 of differentiation. **P < 0.02 (n = 4). D Quantification of TFAP4, MYH2, MYH3, MYOG, CKM, and MYMK mRNAs. *P < 0.05, **P < 0.002, ***P < 0.001 (n = 4). P = 0.064 for MYH2. E Western blot analyses of TFAP4, myosin heavy chain (MHC), CKM, and MYOG proteins. β-Tubulin was analyzed as a loading control. Shown are cropped representative western blots. Full-length western blots are presented in Additional file 13. F Quantitative analysis of western blots. *P < 0.05 (n = 3)