Figure 10

Signaling network up-regulated by constitutively activated EGFRvIII mutation. U87MG glioma cells were transfected with a conditional inducible tetracycline (tet-on) sytem. A EGFRvIII expressing line (row 2) as well as vector control (row 1) was treated with tetracycline (1 ug/ml) for 0, 1, 6, 20, 24 and 48 hours as indicated. Activities of signaling kinases were monitored by Qdot-RPPA. Each sample was serially diluted 1:1 to 1:16 for quantification and spotted in triplicate on the arrays, then probed with total and phospho-specific kinase antibodies. Relative fold changes to time 0 were quantified and illustrated with color under each corresponding array images. Black arrows indicate known canonical signaling pathways; Red dashed arrows indicate the cross-talks between Akt, Src and JNK pathways. p38 and NFkB stay inactive.