Fig. 2.

Effect of Src homology phosphotyrosyl phosphatase 2 (SHP2) silencing on epidermal growth factor (EGF)-induced signaling. Serum-starved control and SHP2-silenced cells were treated with EGF for variable times, and lysates prepared from them were analyzed for Ras, extracellular signal-regulated kinases 1 and 2 (ERK1/2), and Akt activation. a Analysis of Ras activation in the MDA-MB-231 cells. b Analysis of Akt and ERK1/2 activation in the MDA-MB-231 cells. c Analysis of Ras activation in the MDA-MB-468 cells. d Analysis of Akt and ERK1/2 activation in the MDA-MB-468 cells. e Analysis of epidermal growth factor receptor (EGFR) protein level in the indicated cells growing in regular (serum-containing) growth medium. f EGFR band density measurements done using ImageJ software (National Institutes of Health, Bethesda, MD, USA). Data presented are mean ± standard error of the mean of at least three independent experiments. In addition, the EGFR band densities in various lanes were adjusted using β-actin band densities. g Immunoblot analysis of tumor protein extracts for EGFR, phosphorylated ERK1/2 (pERK1/2), phosphorylated Akt (pAkt), and SHP2. GST-RBD glutathione S-transferase-fused Ras-binding domain