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. Author manuscript; available in PMC: 2016 Jan 30.
Published in final edited form as: Oncogene. 2014 Oct 27;34(31):4078–4088. doi: 10.1038/onc.2014.336

Figure 2.

Figure 2

uPAR regulates availability of SH2-binding sites in EGFRvIII. (a) EGFRvIII-expressing and wt-EGFR-over-expressing U87MG cells were transfected with uPAR-specific (+) or NTC (−) siRNA, transferred to SFM, and treated with 10 ng/mL EGF or with vehicle for 10 min. Cell extracts were isolated and incubated with GST-SH2 coupled to glutathione-Sepharose for 3 h at 4°C. The Sepharose beads were wash ed and re-suspended in SDS-sample buffer for SDS-PAGE. EGFR that affinity-precipitated with GST-SH2 was determined by immunoblot analysis. Total cell extracts also were subjected to immunoblot analysis as a control for load. (b) EGFRvIII-expressing U373MG cells were transfected with uPAR-specific (+) or NTC (−) siRNA, transferred to SFM, and treated with 10 ng/mL EGF or with vehicle for 10 min. Binding of EGFRvIII to GST-SH2 was determined as described in panel a.