Fig. 6

TBP inhibits myoblast proliferation and promotes myogenic differentiation. A Relative TBP expression with TBP overexpression in vitro (n = 6). B Relative mRNA levels of several cell cycle genes with overexpression of TBP (n = 4). C The proliferation of transfected CPMs was assessed by 5-ethynyl-2’-deoxyuridine (EdU) incorporation (n = 3). D The proliferation rate of myoblasts after the overexpression of TBP (n = 8). E CCK-8 assays were performed in CPMs with TBP overexpression (n = 6). F Cell cycle analysis of myoblasts after the overexpression of TBP (n = 4). G–I MyHC immunostaining (n = 3) (G), myotube area (%) (n = 8) (H) and myoblast fusion index (n = 8) (I) of CPMs transduced with TBP overexpression. Cells were differentiated for 72 h after transfection. J and K Relative mRNA (n = 6) (J) and protein (n = 3) (K) expression levels of myoblast differentiation marker genes with TBP overexpression. L Relative TBP expression with TBP interference in vitro (n = 3). M Relative mRNA levels of several cell cycle genes with interference of TBP (n = 4). N The proliferation rate of myoblasts after the interference of TBP (n = 8). O The proliferation of transfected CPMs was assessed by 5-ethynyl-2’-deoxyuridine (EdU) incorporation (n = 3). P CCK-8 assays were performed in CPMs with TBP interference (n = 6). Q Cell cycle analysis of myoblasts after the interference of TBP (n = 4). R–T yotube area (%) (n = 8) (R), MyHC immunostaining (n = 3) (S) and myoblast fusion index (n = 8) (T) of CPMs transduced with TBP interference. Cells were differentiated for 72 h after transfection. U and V Relative mRNA (n = 6) (U) and protein (n = 3) (V) expression levels of myoblast differentiation marker genes with TBP interference. In panels K and V, the numbers shown below the bands were folds of band intensities relative to control. Band intensities were quantified by ImageJ and normalized to GAPDH. Data are expressed as a fold-change relative to the control. Results are shown as mean ± SEM. In panels A, B, D–F, H–J, L–N, O–R and T, U), the statistical significance of differences between means was assessed using an independent sample t-test. (*P < 0.05; **P < 0.01; N.S., no significant difference)