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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: Cancer Discov. 2011 Nov 1;1(6):524–538. doi: 10.1158/2159-8290.CD-11-0124

Figure 1. EGFRvIII stimulates mTORC2 activation in vitro and in vivo.

Figure 1

(A) Biochemical analysis of EGFRvIII/EGFR signaling on mTORC2 biomarkers using U87 isogenic cells. Cell lines were cultured in serum-free media for 24 hours.

(B) Immunoblot analysis of LN229 GBM cells in which EGFRvIII was placed under a doxycycline regulatable promoter.

(C) Representative immunohistochemical images demonstrating p-EGFR(Y1068), p-Akt(S473) and p-NDRG1(T346) to assess EGFRvIII-mediated mTORC2 signaling. Scale bar, 20 μm.

(D) Effect of PTEN reconstitution on EGFRvIII-mediated mTORC2 signaling.

(E) Lysates form U87 isogenic cells were subjected to immunoprecipitation with either Rictor antibody or control IgG. Rictor immunoprecipitate was divided into equal fractions for separate kinase reactions using Akt1 purified from insect cells as the substrate. PP242 was added to the kinase reaction in EGFRvIII cells. mTORC2 in vitro kinase activity was assessed by Akt S473 phosphorylation.

(F) The schema of EGFRvIII-stimulated mTORC2 signaling.

See also Supplementary Figure S1.