Fig 1: 17-AAG and gamitrinib increase fractional OXPHOS and restore ETS flux in the presence of ΔGATP.(A) Log2 Abundance of TRAP1 in PBMC and leukemia cells. (B) Comparison of OXPHOS kinetics in presence of the TRAP1 inhibitor, 17-AAG (15 µM); n = 4 independent cell experiments. Comparison of (C) Fractional OXPHOS and (D) FCCP Effect in MV-4–11 cells treated with DMSO or 17-AAG (15 µM). (E) Comparison of respiratory flux inhibition within MV-4–11 cells across a range of ΔGATP in the absence and presence of 17-AAG (15 µM). Respiration was stimulated by the addition of FCCP (1 µM), followed by PCR titration to manipulate ΔGATP. (F) Relative abundance of TRAP1 mRNA following shRNA knockdown of TRAP1 in MV-4–11 cells. (G) Basal respiration in intact cells. (H) OXPHOS kinetics in permeabilized MV-4–11 cells infected with lentivirus encoding shRNA targeted to TRAP1 (TRAP1 KD) or scrambled shRNA (Control). (I–J) Fractional OXPHOS and FCCP Effect measured in the presence of vehicle (DMSO), 17-AAG (15 µM), or gamitrinib (1 µM) in control and TRAP1 KD cells. (K) OXPHOS kinetics in permeabilized MV-4–11 cells in the presence of DMSO or curcumin (10–20 µM). FCCP Effect is graphed to the right. (L) Schematic depicting the presumed mechanism of action of 17-AAG, gamitrinib and curcumin in which the compounds selectively block ATP uptake via the VDAC-ANT axis to restore OXPHOS kinetics. (M) Cell proliferation expressed as a percentage of Control. (N) Cell viability in MV-4–11 cells infected with lentivirus encoding shRNA targeted to TRAP1 or scrambled shRNA and treated for 24 hr with increasing concentrations of Ara-C. Additional treatments included gamitrinib (1 µM), 17-AAG (15 µM) or curcumin (10–20 µM) either alone or plus Ara-C (20 µM). Data depicted as viability based on the percentage of vehicle using the propidium iodide assay. (A) n = 4/6/group; (B–D) n = 4 independent experiments; (E–K) n = 3 independent experiments; (M) n = 8 independent experiments; (N) n = 4–6 independent experiments. Data are presented as mean ± SEM analyzed by unpaired t-tests (F–H, M) two-way ANOVA (B, K), one-way ANOVA (A, I, J, N), paired t-tests (B–E). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.Figure 6—source data 1.Raw values for 'Figure 6' and 'Figure 6—figure supplement 1'.