Extended Data Fig. 3: Corresponding glycine substitutions in BRAF and CRAF confer belvarafenib resistance.
From: ARAF mutations confer resistance to the RAF inhibitor belvarafenib in melanoma
a, Homology model of the ARAF kinase domain (KD) bound to belvarafenib based on the BRAFKD–belvarafenib co-crystal structure. Highlighted residues are mutated in ARAF in belvarafenib-resistant cells. b, Confirmation of BRAF- and CRAF-mutant overexpression in doxycycline-inducible IPC-298 cell lines treated with doxycycline, assayed by Flag–BRAF or Flag–CRAF western blot. c, Cellular viability of doxycycline-inducible IPC-298 cells expressing wild-type BRAF, BRAF(G534D), wild-type CRAF, or CRAF(G426D) and treated with belvarafenib for 3 days ± doxycycline. Data are mean ± s.e.m., n = 3 replicates. d, MAPK signalling in doxycycline-inducible IPC-298 cells expressing wild-type BRAF, BRAF(G534D), wild-type CRAF, or CRAF(G426D) and treated with 0.1, 1 or 10 μM belvarafenib for 24 h. e, Dimer interface of BRAF (PDB code 4MNF). E586 from each BRAF monomer (yellow and cyan) forms hydrogen bonds across the dimer interface with E586 and T589 from the interacting protomer in trans. E586 and T589 are shown as stick models, G534 is shown as spheres. f, MAPK signalling in IPC-298 cells transiently transfected with ARAF(G387D), ARAF(G387D/R362H), ARAF(G387D/E439K), ARAF(G387D/K336M), wild-type ARAF or empty vector (EV). After 24 h, cells were treated with 0.1, 1 or 10 μM belvarafenib for 24 h. g, MAPK signalling in IPC-298 and BRC9 cells after 24-h treatment with 1 μM belvarafenib.
