Fig. 1: ARF6 is sufficient to control oncogenic BRAF protein levels through protein translation.
From: Rapid activation of ARF6 after RAF inhibition augments BRAFV600E and promotes therapy resistance
a Relative amount of MAPK signaling proteins in tumor cells derived from BrafV600E; Cdkn2af/f; Arf6WT or BrafV600E; Cdkn2af/f; Arf6Q67L mice, detected by Reverse Phase Protein Array, two-tailed t-test. n = 3 replicates per cell line. b–h, j, k Western Blot for indicated proteins. b Murine melanoma cells derived from BrafV600E; Cdkn2af/f; Arf6WT, or BrafV600E; Cdkn2af/f; Arf6Q67L mice., n = 3 biological independent experiments. c Human A375 melanoma with doxycycline (DOX)-inducible ectopic expression of ARF6Q67L, BRAFV600E Western blot n = 3 biological independent experiments, h = hours. d Human UACC.62 and A375 cells with or without adenoviral-mediated ectopic expression of ARF6Q67L, control= empty vector. e 4 μM QS11 for 48 h in human A2058 melanoma, HT-29 colorectal carcinoma, and DBTRG-05MG glioma. 2 μM QS11 for 24 h in other cell lines. f 2 μM QS11, h = hours. g 17-AAG and QS11 for 24 h in A375 cells. h 20 μg/ml cycloheximide (CHX) in A375 cells with doxycycline (Dox)-inducible ectopic expressed ARF6Q67L. BRAFV600E protein quantification at 48 h., n = 3 biological independent experiments. i Quantitative RT-PCR for BRAF mRNA in A375 cells with doxycycline (Dox)-inducible ectopic expressed ARF6Q67L, n = 3 biological independent experiments, h=hours. j 4 μM QS11 and 250 nM Torin 1 in A375 cells. BRAFV600E protein quantification at 48 h, n = 3 biological independent experiments. k A375 cells with doxycycline (Dox)-inducible ectopic expressed ARF6Q67L. h = hours. b, c, h, j Two-tailed ratio paired t-test.
