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. Author manuscript; available in PMC: 2007 Aug 1.
Published in final edited form as: Mol Cell. 2006 Aug;23(4):561–574. doi: 10.1016/j.molcel.2006.07.015

Figure 5.

Figure 5

pRKIP Co-Localizes with Activated Raf-1, and Raf-1 Depletion or MEK Inhibition Reverses the Effect of RKIP Depletion on Mitotic Index(A) Localization of pS338 Raf-1 (rhodamine) and pRKIP (FITC) in prometaphase, metaphase, and anaphase/telophase Ptk-1 cells. Scale bar, 20 μm. (B) RKIP-depleted and control HeLa cells transfected with Raf-1, B-Raf, or control siRNA for 48 hrs, were immunoblotted for Raf-1 or B-Raf and mitotic cells counted. (C) Starved RKIP-depleted or control HeLa cells were untreated or treated with 100 ng/ml EGF for 5 min and immunoblotted for pERK and pMEK. (D) After synchronization RKIP-depleted or control HeLa cells were untreated or treated with 10 μM PD098059 for 4 hrs and mitotic cells counted. (E) RKIP-depleted and control HeLa cells were infected with dnMEK1 lentivirus or control lentivirus and mitotic cells counted. (F) RKIP-depleted HeLa cells were treated with 10 μM U0126, or infected with dnMEK1 lentivirus or control lentivirus, and stimulated with EGF as in (C). Lysates were immunoblotted for pERK. α-tubulin was the loading control (B, C, F). All error bars are +/-SE.