E2Fs promote glycolysis in BEC-organoids.
(A-B) Seahorse Substrate Oxidation Assay using UK5099, a mitochondrial pyruvate carrier inhibitor (A), and assessment of the glucose dependency (B) in CD-derived BEC organoids. n=7 for control (Ctrl), n=8 for UK5099. (C) Scheme depicting the treatment of CD/HFD-derived BEC-organoids with FA mix. (D-E) Proton efflux rate (PER) (D), and basal and compensatory glycolysis (E) measured using Seahorse XF Glycolytic Rate Assay. Relative to panel C. n=13. (F) Scheme depicting the treatment of CD-derived BEC-organoids with E2F inhibitor, HLM006474. (G) RT-qPCR of selected cell cycle and glycolytic genes, relative to panel H. n=4. (H-I) PER during the Seahorse XF Glycolytic Rate Assay (H), and basal and compensatory glycolysis (I), relative to panel F. n=10 for control (Ctrl), n=11 for HLM006474.
(J-K) PER during the Seahorse XF Glycolytic Rate Assay (J), and basal and compensatory glycolysis (K), relative to panel C and treatment with HLM006474. n=12 for control (Ctrl), n=11 for HLM006474.
Data are shown as mean ± SD (SEM for A, D, H, J). Absence of stars or ns, not significant (p > 0.05); *p < 0.05; **p < 0.01; ***p <0.001; ****p < 0.0001; unpaired, two-tailed Student’s t-test (G), two-way ANOVA with Sidak’s test (B, E, I, K) were used.
The following figure supplements are available for figure 4:
Figure supplement 1. E2F activation correlates with increased glycolysis.