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. 2007 Mar;18(3):1073–1082. doi: 10.1091/mbc.E06-05-0406

Figure 7.

Figure 7.

Overexpression of GLS-EGFP and ELS-EGFP inhibits mTORC2. (A) Overexpression of GLS-EGFP and ELS-EGFP disrupts normal actin organization and inhibits normal cell spreading. EGFP, GLS-EGFP, and ELS-EGFP were transiently expressed in HeLa cells. Actin cytoskeleton organization was determined by phalloidin-TRITC staining. The arrowheads indicate transfected cells. (B) Overexpression of GLS-EGFP and ELS-EGFP inhibits cell spreading. The level of GFP signal in individual cells was categorized as in Figure 6B. (C) ELS and GLS do not inhibit AKT/PKB phosphorylation in HEK293 cells. ELS-EGFP, GLS-EGFP, or EGFP was transiently expressed with GST-AKT in HEK293 cells. The cells were starved from fetal bovine serum for 4 h before incubated with insulin for 15 min. Phosphorylation of GST-AKT and endogenous AKT was analyzed by Western blot with a P-Ser473 antibody. (D) ELS and GLS do not inhibit AKT/PKB phosphorylation in HEK293T cells. ELS-EGFP, GLS-EGFP, or EGFP was transiently expressed with GST-AKT in HEK293T cells. Phosphorylation of GST-AKT and endogenous AKT was analyzed by Western blot with a P-Ser473 antibody.