Fig. 7.

The effect of eliminating ZBP-89 expression via ZBP-89 siRNA on the C2C12 myogenic program. (A) MBs (5×10⁴) were incubated in GM, DM or transfected with either si-ZBP-89 or siControl in GM then visualized with a light microscope for 18 and 36 h as described in Fig. 1B. (B) qPCR analysis of endogenous ZBP-89 mRNA levels normalized to U6 expression isolated at 48 h as described in Fig. 3A (C) qPCR analysis of endogenous myogenin mRNA levels normalized to U6 expression isolated at 48 h as prepared and described in panel B. (D) Western blots analysis of WCEs (50 μg) isolated from C2C12 cells as grown in panel A for 96 h as described in Materials and methods. β-tubulin serves as a loading control. (E) MT cells (5×10⁴) were allowed to differentiate for 24 h and then transfected with either siZBP-89 or siControl. Cells were harvested 48 h later and qPCR analysis was performed as mentioned in panel B. Experiments were performed in triplicate three times with bars representing the standard error of the mean.