Fig. 2

ARHI induction increases glucose uptake and glycolytic activity. a Immunofluorescent staining of SKOv3-ARHI cells for GLUT1 (green) at 24 and 48 h post-induction with doxycycline (DOX). The nuclei are stained with DAPI (blue) for reference. b Immunofluorescent staining of SKOv3-ARHI cells for GLUT1 and ARHI at 48 h post-induction. c SKOv3-ARHI cells were induced with Dox at the given time points and the uptake of [³H]-2-deoxyglucose determined. SKOv3-ARHI cells were stably transfected with shRNA for Atg5 to block the initiation of autophagy (shATG5). Uptake was measured as cpm/10⁵ viable cells after 30 min of incubation. Three independent samples for each condition were used (n = 3) to compute the mean and standard deviation and the statistical difference in uptake between samples was determined by unpaired two-tailed t test (*, p < 0.05. **, p < 0.01). d SKOv3-ARHI or e HEY-ARHI cells were treated with 2-deoxglucose (2-DG) and induced with Dox in media for the specified times (n = 3 for each condition). Percent cell viability was determined by SRB assay. Statistical difference was determined by unpaired two-tailed t test (*, p < 0.05; ****, p < 0.0001). f Normalized extracellular lactate signal measured by ¹³C-NMR between 24 and 32 h of ARHI induction autophagy. Three independent samples were for each condition were used to calculate the mean and standard deviation and statistical significance was determined by two-way ANOVA with p-values adjusted for multiple comparisons (***, p < 0.001)