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. 2015 Dec 2;5:17626. doi: 10.1038/srep17626

Figure 2. Fumos inhibits Shp2-dependent signaling pathways.

Figure 2

(a) Fumos inhibits EGF-induced ERK1/2 activation. Serum starved cells were pretreaed with various concentrations (0, 10, 20, 30, 40 μM) of Fumos for 24 h and then stimulated with EGF (20 ng/ml, 5 min). The control treatment is not induced by EGF. (b) Fumos doesn’t inhibit PMA-induced ERK1/2 activation. Cells were pretreated with increasing concentrations of Fumos for 24 h, and then stimulated with 100 nM PMA for 5 min. DMSO treatment as control. ERK1/2 activation was analyzed by Western-Blot with antibodies to phosphorylated ERK1/2 (pERK) or ERK1/2. GAPDH is used as a loading control for Western Blot. The control treatment is not induced by PMA. (c) Fumos inhibits EGF-induced Ras activation. Serum-starved cells were treated with Fumos(20 μM) and EGF as indicated. Cell lysates (0.5 mg of protein/each) were incubated with GST-RBD-agarose to pull down active-Ras-GTP, which was visualized by immunoblotting with an anti-Ras antibody. (d) Fumos doesn’t inhibit RasV12-induced ERK1/2 activation. HEK 293T cells were transiently transfected with a pCMV vector for RasV12 and empty vector, and treated with Fumos(20 μM) or U0126(10 μM) for 24 h, respectively. Cell lysates were immunoblotted with Myc-tag indicating the expression of RasV12. Total cell lysates were immunoblotted with antibodies for p-ERK1/2 and ERK1/2.