Fig. 7: VARS controls the FA metabolism in resistant melanoma cells.
From: Valine aminoacyl-tRNA synthetase promotes therapy resistance in melanoma
a, Experimental set-up showing the score attribution for VARS signature (high and low) in patients with BRAFV600E-mutated SKCM melanoma from TCGA (n = 233). GSEA analysis enrichment of the FA metabolism in VARS high versus VARS low TCGA melanomas. b, FA oxidation (FAO) activity measured in SENS and RES M395 patient-derived cultures (mean fold change ± s.e.m.; n = 3 independent biological replicates, two-tailed unpaired t-test). c, Cell death measurement by fluorescence staining with PI by FACS in M395 RES cells treated or not with etomoxir and vemurafenib at indicated concentrations (mean fold change ± s.e.m.; n = 3 independent biological replicates; two-way analysis of variance (P < 0.0001)). Tukey’s multiple comparisons test are indicated. d–f, Western blot of VARS and HADH protein expression in SENS and RES M395 cells (d), in A375, M395 and MM029 RES cells depleted of VARS (e), and in A375, M395 and MM029 SENS cells overexpressing (VARS) or not (control) VARS (f). g,h, FAO activity measured in M395 RES cells depleted or not for VARS (g) and in M395 SENS cells overexpressing (VARS) or not (control) (h) (fold mean ± s.e.m. of n = 3 independent replicates, two-tailed unpaired t-test). i, FAO activity measured in M395 and MM029 RES cells depleted of HADH (shHADH-1 and shHADH-2) compared with control cells (shCTRL) (mean fold change ± s.e.m.; n = 2 M395 RES and n = 3 MM029 RES, two-tailed unpaired t-test). j, Cell death measurement by fluorescence staining with PI by FACS in control (shCTRL) or HADH (shHADH) depleted M395 RES cells treated or not with vemurafenib (as indicated) (mean fold change ± s.e.m.; n = 3 independent biological replicates, two-way analysis of variance (P < 0.0001). Tukey’s multiple comparison tests are indicated.
