Figure 5 – Glucose uptake capacity is sufficient to drive OXPHOS inhibitor resistance.
A: Increase in OP-ind responses stimulated by insulin. Histograms show AMPKAR2Δ responses to 1.8 μg/ml oligomycin. N=2.
B: Change in OP-dep responses in response to inhibitor treatment for MCF10A cells under 17 mM glucose. Inhibitors were added 30 minutes prior to oligomycin. Horizontal black lines indicate the fraction of OP-dep cells under control treatment (DMSO); points falling outside the gray region are considered significant by t-test. Points represent the mean, and error bars standard error of the mean; N=2.
C, D and E: Effect of increased glucose uptake on OXPHOS inhibitor responses. MCF10A-AMPKAR2 cells stably overexpressing GLUT1-IRES-NLS-mCherry were cultured with 0 (C), 100 (D), or 10000 ng/ml (E) insulin and exposed to oligomycin (OM). Each row in the heatmaps (upper panels) represents an individual cell; rows are sorted by relative mCherry intensity (corresponding to the level of GLUT1 overexpression), which is indicated by the color bar to the left. GLUT1 expression levels are normalized to the minimum and maximum expression levels in the population. Lower panels show scatter plots of mCherry intensity and AMPKAR2Δ following oligomycin treatment. N=2.
F, G: Increase in OP-ind responses following glucose starvation. (F) shows average AMPKAR2PHOS recordings for MCF10A cells grown in the absence of glucose for 24 hours and then treated with glucose at the specified concentrations, followed by 1.8 μg/ml oligomycin at 30 minutes or 1 minute after glucose addition. (G) shows histograms of AMPKAR2Δ values after oligomycin treatment for the conditions shown in (F). N=2.