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. 2015 Dec 8;291(5):2181–2195. doi: 10.1074/jbc.M115.676510

FIGURE 6.

FIGURE 6.

AA activates the MEK-ERK1/2 pathway and promotes C2C12 cell proliferation independent of Ras/Raf. a, levels of Ras, t-Raf (c-Raf), and p-Raf (p-c-Raf) proteins in C2C12 cells in response to AA (5 mm) treatment were assessed by Western blotting at the indicated times. b, normalized level of Ras and p-Raf. Ras protein was normalized to β-actin, and p-Raf was normalized to total Raf. c, protein levels of Ras, t-Raf/p-Raf, t-MEK/p-MEK, and t-ERK1/2/p-ERK1/2 were determined by Western blot analysis in normal C2C12 and DN-Ras C2C12 cells treated with AA (5 mm) or PBS for 12 h. d, quantified levels of p-MEK and p-ERK relative to t-MEK and t-ERK, respectively, based on blots in c. e, protein levels of Ras, p-Raf, t-Raf, p-MEK, t-MEK, p-ERK, t-ERK, and β-actin in C2C12 cells treated with AA (5 mm) for 12 h in the presence or absence of the inhibitors FTA or GW5074 (GW) were determined by Western blotting. f, cell proliferation was analyzed in C2C12 cells or DN-Ras expressing C2C12 cells treated with 5 mm AA. g, cell proliferation was analyzed in C2C12 cells treated with AA (5 mm) in the presence or absence of FTA or GW5074. h, schematic model of AA function. Similar results were obtained in three separate experiments. Data are presented as means ± S.E. (error bars; *, p < 0.05; ***, p < 0.001).