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. 2011 Oct 31;195(3):377–386. doi: 10.1083/jcb.201106138

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© 2011 Hwang et al.

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Figure 2. — CIIA regulates EGF-induced Ras and Erk activation. (A) MDCK/control or MDCK/CIIA-Flag cells were deprived of serum for 16 h and incubated for 5 min with or without 100 ng/ml EGF. Cell lysates were immunoprecipitated (IP) with anti-SOS1 antibody, and the pellets were immunoblotted (IB) with anti-Ras antibody. Cell lysates were also examined directly by immunoblotting with antibodies to SOS1, Ras, or Flag. (B and C) MDCK/CIIA-Flag and MDCK/control cells (B) or HeLa cells stably expressing either GFP (control) or CIIA siRNA (C) were deprived of serum for 16 h and then incubated for 5 min with or without 100 ng/ml EGF. Cell lysates were subjected to pull-down (PD) with GST-RBD and assayed for Ras activity.