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. 2011 May 26;286(29):25903–25921. doi: 10.1074/jbc.M111.243030

FIGURE 5.

FIGURE 5.

GRAF promotes terminal differentiation of myoblasts in a GAP- and SH3-dependent fashion. A, schematic showing full-length FLAG-tagged GRAF (F-GRAF) and GRAF variants used herein. PH, pleckstrin homology; S/P, serine/proline-rich domain. B and C, C2C12 cells were transfected with FLAG GRAF1 (F-GRAF) or FLAG GRAF1R412Q (F-GAPm) and exposed to LSM for 48 (B) or 72 h (C). Cells were stained with anti-FLAG antibody to detect GRAF variants (green) and co-stained with phalloidin and tropomyosin (Tm) or p21cip. Scale bars, 50 μm. D, quantification of GRAF-induced skeletal differentiation marker gene expression. The number of cells positive for the indicated marker was counted in control (Con) and GRAF (or indicated GRAF variant-)-expressing cells exposed to LSM for 48 h. Data represent the % of expressing cells counted in three to five separate experiments (n = 200–350 cells/condition).