Supplementary Figure 4: Additional characterization of in vito Mpc1 Lgr5-KO.

(a) Flow cytometry profile for organoids from control and MPC1 knockout cultures showing GFP positivity. (b) Long-term cultures retain increased growth ability throughout extended passaging (over ∼2 months) (n = 15 control and knockout, scale bar = 200 μm) (c) Control and MPC1 knockout organoid western blots for MPC1 and MPC2 and pyruvate dehydrogenase (phospho 293 and total). (d) Transcript abundance of MPC1 knockout organoids showing stem and differentiation marks (n = 3). (e) Quantification of specific Acetyl Histone H3 marks in control and knockout organoids (paired control set to 1.0) (n = 8 organoids). (f) Steady state metabolite abundance for control and MPC1 knockout organoids (n = 6 control and knockout). Data are mean ± s.e.m. ^∗P < 0.05, ^∗∗P < 0.01, ^∗∗∗P < 0.001. All p values were calculated using Student’s t-test. Unprocessed western blots are provided in Supplementary Figure 7.