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. Author manuscript; available in PMC: 2013 Apr 13.
Published in final edited form as: Cell. 2012 Apr 13;149(2):307–321. doi: 10.1016/j.cell.2012.02.053

Figure 6. AZD6244-mediated kinome reprogramming in C3Tag mouse model of TNBC.

Figure 6

(A) PDGFRβ is induced in C3Tag tumors after 2d AZD6244 treatment, as shown by anti-PDGFRβ immunofluorescence.

(B) AZD6244 treatment of C3Tag mice for 2 and 7d causes c-Myc degradation and induced PDGFRβ expression, as shown by western blot.

(C) Tumor-derived C3Tag cell line shows AZD6244-mediated c-Myc loss and induction of RTKs. T2-C3Tag cells were treated with AZD6244 and RTK reprogramming determined by western blot.

(D) Expression of c-Myc(T58A) in T2-C3Tag cells suppresses AZD6244-mediated RTK reprogramming. T2-C3Tag cells stably expressing vector or human c-Myc(T58A) were treated with AZD6244 for 24h and analyzed by western blot.

(E) MIB/MS profile of C3Tag tumors in response to AZD6244 for 28d, sorafenib for 26d, or combined AZD6244 and sorafenib for 26d show distinct kinome responses. The line graphs show iTRAQ-determined quantitative changes in MIB binding as a ratio of inhibitor/untreated. Ratio <1 denotes decreased MIB binding and >1 increased MIB binding of kinases in treated versus control tumors.

(F) AZD6244 plus sorafenib inhibits AZD6244-induced kinome response in C3Tag tumors, as identified by MIB/MS. Changes in MIB binding (>1.5 fold) of AZD6244-responsive kinases following cotreatment with sorafenib are shown in yellow.

(G) Cotreatment of C3Tag mice with AZD6244 and sorafenib prevents upregulation of kinases observed by AZD6244 treatment alone, including CDKs and PDGFRβ. These kinases were selected from (E) with a >1.5-fold reduction in MIB binding between AZD6244 treatment alone and cotreatment with AZD6244 and sorafenib.

See also Figure S6.