Quantification of efficiency of Gls and Mpc2 deletion. (A) Raw chimerism frequencies for Gls Mpc2 chimeras in Fig. 8 B. Each symbol represents an individual mouse. Mean values ± SEM are shown. Data are pooled from two independent experiments. ***, P < 0.001; ****, P < 0.0001 by two-way ANOVA with post-hoc Tukey’s multiple comparisons test. (B) Quantitative PCR analysis of genomic Gls and Mpc2 deletion in recent Gls Mpc2 KO chimeras. CD45.2⁺ cells from the bone marrow or spleen were sorted for genomic DNA extraction. DNA quantification within loxP sites was quantified relative to WT cells and GAPDH. Mean values ± SEM are shown. Data are pooled from two independent experiments. ****, P < 0.0001 by Student’s two-tailed t test. (C) RNA-seq reads across exons 1–3 of Gls (top) and 2–5 of Mpc2 (bottom) of CD45.2⁺ GMPs from ROSA26 CreER^+/− (WT), Mpc2 KO, Gls KO, or Gls Mpc2 DKO chimeras about 10 wk after tamoxifen treatment. Exon 1 of Gls and exon 3 of Mpc2 are floxed in the appropriate genotyped mice. One representative trace for each genotype is shown using Integrative Genomics Viewer (left). TPMs were quantified for the floxed exon of Gls (top right) or Mpc2 (bottom right). TPMs were calculated from reads obtained using DEXSeq. Each data point represents a mouse (n = 5 for each genotype). Mean values ± SEM are shown. ***, P < 0.001; ****, P < 0.0001 by one-way ANOVA with post-hoc Tukey’s multiple comparisons test.