Fig. 4.

SLC25A1 affects mitochondrial activity and self-renewal in CSCs. a Oxygen consumption rate (OCR) were assessed using the Seahorse Extracellular Flux Analyzer after injection of oligomycin (O), FCCP (F) and Antimycin/Rotenone (A + R), in H1299 cells expressing SLC25A1 grown as monolayer or spheres. OCR was normalized to cell number. To eliminate the influence of the media composition and of attachment, for all these experiments monolayer and sphere cultures were first grown for several days, and subsequently they were dissociated, plated on 96-wells coated with the geltrex ECM and pre-equilibrated in the same media containing serum and 5% glucose for 6–12 h, prior to the assessment of the OCR. BR, MR, SRC refer to basal, maximal respiration and spare respiratory capacity. b OCR rates in control or in cells expressing SLC25A1 shRNAs. c OCR rates of the indicated tumor cells in vehicle (black bars) or CTPI-2 treated (gray bars) cells. In all cases, treatment with CTPI-2 was for 3 h and OCR levels were normalized to cell number. There were no changes in viability during this period of time. Bars represent the standard deviation, asterisks refer to **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001 by unpaired T-test