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. Author manuscript; available in PMC: 2013 Aug 23.
Published in final edited form as: Mol Cell. 2011 Apr 8;42(1):50–61. doi: 10.1016/j.molcel.2011.03.017

Figure 3. Rac1 Regulates mTOR Activity Independently of PI3K, Binding to GTP/GDP and Actin Polymerization.

Figure 3

(A) Primary Rac1 fl/fl MEFs were grown in 10% serum and treated or not with 4-HT for 48 hr, then serum starved (overnight). They were incubated or not with 100 nM wortmannin for 30 min then subjected to stimulation with 1 μM PMA for 15 min.

(B) Cells were grown in 10% serum and treated with ethanol (control) or with 500 nM 4-HT for 48 hr to delete Rac1. In some conditions, cells were treated for 1 hr with 100 nM wortmannin or 100 nM Rapamycin at 37°C or with 10 μM of a Rac inhibitor, NSC-23766.

(C) MEFs were treated or not with 500 nM 4-HT for 36 hr in DMEM with 10% heat-inactivated FBS to delete Rac1. Cells were then infected with recombinant vaccinia viruses, expressing Rac1 WT, V12Rac1, N17Rac1, or the empty vector pSC65 for 14 hr.

(D) HeLa cells were grown in 10% serum then serum starved for 14 hr. They were treated or not with 10 μg/ml cytochalasin D and stimulated or not with 10% serum supplemented with 50 nM PDGF. Cells were then lysed and subjected to SDS-PAGE and western blot analysis. These results are representative of three experiments.