In vitro GMPs do not require the MPC for survival. (A) WT GMPs were cultured with DMSO or 10 μM UK5099 for 7 d. On each day of the culture, cells were harvested, and live cells were counted. One experiment with 10 technical replicates from two mice per group is depicted as a representative of four independent experiments. P values >0.05 by two-way ANOVA with post-hoc Tukey’s multiple comparisons test are not depicted. (B) Pyruvate excretion rate in the media of GMPs cultured with DMSO or UK5099 over a 48-h culture. ****, P < 0.0001 by Student’s two-tailed t test. Mean values ± SEM are shown for four replicates from two independent experiments. (C) GMPs from Mpc2^fl/fl; ROSA26 CreER^+/− or ROSA26 CreER^+/− mice were treated with 0.5 μM 4-OHT. On each day of the culture, cells were harvested, and live cells were counted. One experiment with 10 technical replicates from two mice per group is depicted as a representative of two independent experiments. P values >0.05 by Student’s two-tailed t test are not depicted. (D) Quantitative PCR analysis of genomic Mpc2 deletion in GMPs from Mpc2^fl/fl; ROSA26 CreER^+/− or ROSA26 CreER^+/− mice. GMPs were treated with 4-OHT for 2 d prior to DNA extraction. DNA quantification within loxP sites was quantified relative to WT cells and GAPDH. Mean values ± SEM are shown for four replicates from two independent experiments. ****, P < 0.0001 by one-way ANOVA with post-hoc Tukey’s multiple comparisons test.