Oncogenic AKAP9-BRAF fusion is a novel mechanism of MAPK pathway activation in thyroid cancer
J. Clin. Invest. Raffaele Ciampi, et al. 115:94 doi:10.1172/JCI23237 [Go to this article.]

Figure 4
The AKAP9-BRAF chimeric protein has increased BRAF kinase activity and results in NIH3T3 cell transformation. (A) In vitro kinase activity of myc-tagged BRAF^WT, AKAP9-BRAF, and BRAF^V600E in the absence or presence of H-RAS^G12V in COS7 cells. BRAF kinase activity was measured in myc-IgG immunoprecipitates using MEK1 substrate phosphorylation as a read-out. Each sample was assayed in triplicate, and bars represent the standard deviations from the mean. Similar results were obtained in 2 independent transfections. (B) Western blots of lysate from cells stably transfected with pEFP, pEFP-BRAF^WT, pEFP–AKAP9-BRAF, pEFP-BRAF^V600E, or pBabe puro–H-RAS^G12V probed with antibodies to phospho-ERK1/2 (pERK), total ERK (tERK), or C-terminus of BRAF. Similar results were obtained in 3 additional independent experiments. (C) Foci formation of NIH3T3 cells transfected with pEFP-BRAF^WT, pEFP–AKAP9-BRAF, pEFP-BRAF^V600E, or pBabe puro–H-RAS^G12V. Bars represent the average number of foci per plate from 6 plates. Similar results were obtained in 2 independent experiments.