Fig. 4: ARF6 activation protects against MAPKi-induced apoptosis and promotes the development of MAPKi-resistance cells.
From: Rapid activation of ARF6 after RAF inhibition augments BRAFV600E and promotes therapy resistance
a, b Total ARF6 and ARF6-GTP pulldown in A375, 5 μM vemurafenib for 4 h or as indicated, Dabrafenib treatment for 4 h, PF-07799933 treatment for 2 h in 0% FBS media. c–f Apoptosis detection. One-way ANOVA with multiple comparisons. c 1 μM Vemurafenib, dox-inducible ectopic expressed ARF6WT and ARF6Q67L in A375, apoptosis measured at 48 h, Ctrl= no doxycycline, n = 5 replicates per condition. d 1 μM Vemurafenib, 4 μM QS11 for A375, n = 4 replicates per condition, apoptosis measured at 48 h, 2 μM Vemurafenib, 4 μM QS11 for UACC.62, n = 3 replicates per condition, apoptosis measured at 24 h. e 1.25 μM Dabrafenib, 0.0625 μM Trametinib, dox-inducible ectopic expressed ARF6WT and ARF6Q67L in A375, apoptosis measured at 48 h, Ctrl= no doxycycline, n = 3 replicates per condition. f 1.25 μM Dabrafenib, 0.0625 μM Trametinib, 4 μM QS11, apoptosis measured at 48 h, n = 3 replicates per condition. g Western Blot for indicated proteins. 1 μM Vemurafenib, 4 μM QS11 in A375. 2 μM Vemurafenib, 4 μM QS11 in UACC.62. h, i Colony outgrowth assay in A375. Two-tailed unpaired t-test. n = 4 biological independent experiments. h 1 μM Vemurafenib, 4 μM QS11, for 30 days. i 250 nM Dabrafenib, 12.5 nM Trametinib, 2 μM QS11, 4 μM QS11, for 30 days.
