FIG 9.

Stable overexpression of MUNC enhances RNA of myogenic markers but not that of the corresponding proteins. qRT-PCR expression of MUNC isoforms and myogenic markers following C2C12 transfection with linearized vectors encoding the WT unspliced form of MUNC, an unspliceable form of MUNC with a point mutation preventing RNA splicing, and a spliced form of MUNC. Measurements were performed on proliferating cells (GM) and differentiating cells after 3 days in differentiation medium (DM3). Expression data of unspliced MUNC (A and F), spliced MUNC (B and G), MyoD (C and H), myogenin (D and I), and Myh3 (E and J) RNAs are shown. Data are normalized to GAPDH expression and then normalized again in each panel to the level in vector-transfected cells in GM or DM. Data represent means ± standard errors of the means (n = 3). (K) Western blot analysis showing level of MyoD and MyoG proteins in C2C12 cells overexpressing MUNC in GM. Actin was used as a loading control. (L) The same experiment as shown in panel K except performed in cells after 3 days in DM.