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. 2007 May;18(5):1850–1860. doi: 10.1091/mbc.E06-08-0765

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© 2007 by The American Society for Cell Biology

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Figure 4. — Binding of R-Ras to Rgl2/Rlf. (A) Cos7 cells expressing mCFP-tagged R-Ras-Q87L and the Myc-tagged effector proteins indicated at the bottom of the panel were used for the analysis. In p110α expression, p85α was used for coexpression to stabilize the PI3K heterodimer complex. From the cell lysates, GFP-tagged proteins were immunoprecipitated, and bound proteins were analyzed by immunoblotting with anti-Myc mAb or anti-GFP mAb. (B) Cos7 cells expressing HA-tagged R-Ras mutants and Myc-tagged Rgl2/Rlf were lysed and analyzed as described in A. (C) Endogenous R-Ras protein was immunoprecipitated with anti-R-Ras serum from Cos7 cells expressing the Myc-tagged effector proteins. The immunoprecipitates were analyzed as described in A. Preimmune serum was used as a control.