Extended Data Fig. 4: SLC25A1 and MDH1 contribute to TCA cycle metabolism in embryonic stem cells.
From: A non-canonical tricarboxylic acid cycle underlies cellular identity
a, b, Immunoblot of clonal mouse ES cells in which CRISPR/Cas9-mediated editing was used to target either a non-genic region of chromosome 8 (Ctrl) and Slc25a1 (SLC25A1-1 and SLC25A1-2) (a) or Mdh1 (MDH1-1 and MDH1-2) (b). c, d, Fractional M+1 enrichment of malate (Mal), fumarate (Fum), aspartate (Asp) and citrate (Cit) in control (Ctrl) and Slc25a1-edited ES cells (c) or Mdh1-edited ES cells (d) cultured in medium containing [4-2H]glucose. e, f, Fractional M+2 enrichment of citrate, fumarate, malate and aspartate derived from [U-13C]glucose in control and Slc25a1-edited (e) or Mdh1-edited (f) ES cells. g, Steady-state levels of TCA cycle metabolites in Slc25a1-edited or Mdh1-edited ES cells. Levels are represented as the fold change (expressed in log2) relative to chromosome 8-targeted control cells. Data are mean ± SD, n = 3 independent replicates. Significance was assessed in comparison to control cells by two-way ANOVA (c-f) with Sidak’s multiple comparisons post-test.
