Fig. 5: N-MCJ mimetics attenuate mitochondrial respiration in chemoresistant cancer cells.
From: Mitochondrial ATP fuels ABC transporter-mediated drug efflux in cancer chemoresistance
a Diagram of N-MCJ mimetics. Top diagrams show full length MCJ (not to scale). N-term, N-terminus; TM, transmembrane domain; C-term, C-terminus; TAT, HIV TAT sequences; N-MCJ, first 20 aa of the MCJ N-terminus; mts, mitochondrial targeting sequence; mpp, mitochondrial penetrating peptide; rev, reversed aa sequence. b, c NCI/ADR-RES cells were treated with MITOx20 (25 μM) or Control-20 (25 μM) for 12 h and the analyzed by Seahorse MitoStress Assay for b oxygen consumption rates (OCR) at baseline and in response to sequential injections of oligomycin (O), FCCP (F), and rotenone with antimycin (RA) and for c basal (Control n = 5, MITOx20 n = 4) and ATP-linked (ATP) (Control n = 5, MITOx20 n = 4) relative to Control-20 treated cells. p = 0.0243, 0.001 by unpaired t test. d, e NCI/ADR-RES cells were treated with MITOx30 (25 μM) or Control-30 (25 μM) for 12 h and then analyzed as in (b, c) (n = 4). p = 0.0005, 0.002 by unpaired t test. f NCI/ADR-RES cells were incubated with or without (Vehicle) MITOx20 (25 μM) or MITOx30 (25 μM) for 5 h and then total ATP levels were determined by Luciferase assay (n = 4). p = 0.0001, 0.0001 by one-way ANOVA and Tukey’s multiple comparisons test. g NCI/ADR-RES cells were incubated with or without (Vehicle) Control-20 (25 μM) or Control-30 (25 μM) for 5 h and then total ATP levels were determined by Luciferase assay (n = 3). p = 0.3635, 0.7322 by one-way ANOVA and Tukey’s multiple comparisons test. h NCI/ADR-RES cells were analyzed for OCR at baseline and in response to sequential injections of MITOx30 (25 μM), FCCP, and rotenone with antimycin. i Basal OCR (p = 0.0001, 0.9681 by one-way ANOVA and Tukey’s multiple comparisons test), j and ECAR (p = 0.7301, 0.4473 by one-way ANOVA and Tukey’s multiple comparisons test) in NCI/ADR-RES cells in response to injection of MITOx30 or Control-30 as determined by MitoStress assay (vehicle n = 8, Control-30 n = 6, MITOx30 n = 8). Mean ± SEM is shown for all figures. * denotes p < 0.05 by unpaired t test or one-way ANOVA and Tukey’s multiple comparisons test. Results are representative of at least two experiments.
