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. Author manuscript; available in PMC: 2017 Jun 1.
Published in final edited form as: Cancer Discov. 2016 Apr 21;6(6):601–611. doi: 10.1158/2159-8290.CD-16-0075

Figure 2. EGFR-RAD51 is an oncogenic EGFR alteration.

Figure 2

(A) Ba/F3 lines stably expressing pMSCV (vector only), EGFR-WT, EGFR-L858R or EGFR-RAD51 were subjected to western blot analysis with indicated antibodies. The three distinct EGFR variants were detected at the anticipated molecular weight (MW) of ~150kD. EGFR-RAD51 fusion is detected with both the N-terminal EGFR antibody [EGFR(N)] and with the RAD51 antibody. There is no cross reactivity between wild-type RAD51 protein, which has MW ~35kD, and the EGFR-RAD51 fusion.

(B) Ba/F3 cells transfected with indicated constructs (pMSCV = vector only) were grown in the absence of IL-3 and counted every 24 hours. *** = p < 0.0001.

(C) Ba/F3 expressing EGFR variants were serum starved for 16 hours, treated with 50 ng/mL EGF for 5 minutes, and subjected to western blot analysis with indicated antibodies.

(D) Ribbon diagram and space-filling model of the EGFR-RAD51 kinase domains illustrating the proposed mechanism of activation. Purple = first EGFR kinase domain; green = second EGFR kinase domain; red = first RAD51 partner; blue = second RAD51 partner, yellow asterisks = active sites.