PMC

(A) Schematic diagram describing the components and metrics from Seahorse assays used in the PCA; Bioenergetic Health Index (BHI) calculation is described in methods. (B) Metabolic parameters individually tested for correlation (Pearson) to PLX4720 IC50 of the cells. (C) The first principle component of a linear combination of glycolysis and glycolytic reserve correlates with IC50.

(A) Extracellular pH or acidification rate of parental A2058 and WM164, and their Rho0 derived counterparts; bar plot showing maximum ECAR after adding glucose (10 mM), with t-test between parental and Rho0. (B) Oxygen consumption rate of parental A2058 and WM164, and their Rho0 derived counterparts; bar plot showing maximum OCR after adding FCCP (1 uM), with t-test between parental and Rho0. (C) Log2 normalized proliferation of parental A2058 treated with DMSO or 32 uM PLX4720, and Rho0 A2058 treated with DMSO or 32 uM PLX4720 (purple). Similar population doublings-time plot of WM164 (lower panel), with additional dose of 1 uM PLX4720. (D,E) Quantification of percent decrease in Doublings from respective DMSO control (ie., parental compared to parental, or Rho0 compared to Rho0).

(A) Proliferative spectrum of IC50’s for PLX4720 based on DIP rate. The dose-response curves are generated using a 2-fold dilution of PLX4720 from 32 uM down to zero (DMSO). The proliferative rates are calculated using the slope of the log2-normalized population curve after 48 hours (see Methods). (B) Oxygen consumption profiles for the cell lines with sequential additions of oligomycin (1 uM), FCCP (1 uM) and Rotenone/Antimycin A (0.5 uM). (C) Extracellular pH profiles for the cell lines with sequential additions of glucose (10 mM), oligomycin (1 uM), and 2-deoxyglucose (0.5 uM).

(A) Extracellular pH or acidification rate of parental A2058 and WM164, and their ddC/zalcitabine treated counterparts (40 uM zalcitabine); bar plot showing maximum ECAR after adding glucose (10 mM), with t-test between parental and ddC treated cells. (B) Oxygen consumption rate of parental A2058 and WM164, and their ddC/zalcitabine treated counterparts (40 uM zalcitabine); bar plot showing maximum OCR after adding FCCP (1 uM), with t-test between parental and ddC treated cells. (C) Log2 normalized proliferation of ddC treated WM164 (top panel) or ddC treated A2050 (lower panel). (D) Quantification of percent decrease in Doublings from respective DMSO controls for WM164 and A2058 cell line (not statistically significant). (E) Drug Induced Proliferative (DIP) metric dose response curves (non-normalized) marking model estimated EC50 values in nM (red dashed vertical line), and model estimated Emax values (grey dashed horizontal line).