Fig. 2: Ranolazine reduces mitochondrial respiration in BRAFi-resistant cells.
a, Volcano plot showing the log2 fold change of lipids in A375 versus A375VR cells (LPC: lysophosphatidylcholine; MG, monoacylglycerols; DG, diacylglycerols; TG, triacylglycerols; PE, phosphatidylethanolamines and ChE, cholesterol esters). b, Total levels (integrated peak areas; a.u., arbitrary units) of lipids divided by classes in A375 versus A375VR cells. Data are presented as the mean ± s.e.m. based on n = 4 biological replicates analysed by two-tailed unpaired t-test. ***P < 0.001. c, Representative oxygen consumption rate (OCR) of A375 and A375VR cells. Oligo, oligomycin; FCCP, carbonyl cyanide-p-trifluoromethoxyphenylhdrazone; R, rotenone; A, antimycin A. Data are shown as the mean ± s.d. with n = 3 replicate wells. d, Basal OCR of A375VR and A375 cells was plotted against their basal extracellular acidification rate (ECAR). Data are from n = 3 biological replicates of three replicate wells. e, Basal or maximal respiration and ATP production in A375 and A375VR cells. Data are presented as the mean ± s.d. based on n = 3 replicate wells analysed by two-tailed unpaired t-test. **P < 0.01; ***P < 0.001. f,g Representative OCR (f) and basal or maximal respiration and ATP production (g) of A375 and A375VR cells in the absence (DMSO) or presence of 10 µM RANO. Data are shown as the mean ± s.d. based on n = 3 replicate wells and analysed by one-way ANOVA with uncorrected Fisher’s least significant difference (LSD). *P < 0.05; **P < 0.01. h,i Relative cell number of A375 or A375VR cells (h) treated daily with 10 µM or 25 µM RANO or treated once per week with 100 µM RANO with or without 0.5 µM BRAFi (i) as indicated. Control cells were treated with DMSO. Data are presented as the mean ± s.e.m. based on n = 3 biological replicates and analysed by one-way ANOVA with Sidak’s multiple-comparisons test. ***P < 0.001.
