Skip to main content
. Author manuscript; available in PMC: 2022 Sep 1.
Published in final edited form as: Nature. 2021 Aug 18;597(7875):263–267. doi: 10.1038/s41586-021-03827-2

Extended Data Fig. 9. PKM2 ablation in villi results in PKM1 upregulation.

Extended Data Fig. 9.

(A) Representative intestines from 12-week-old mice examined by IHC for the indicated targets (scale bar = 200μm). (B) Mouse intestinal epithelial cell lysates from wild-type mice (WT), VillinCre;PKM2fl/fl mice, and WT mice treated with TEPP-46 were analyzed via enzymatic assay for pyruvate kinase activity (mice per group, left to right: 5|10|5; same final protein concentration in each reaction well). (C) WT and VillinCre;PKM2fl/fl mice were sacrificed and intestines were fixed and examined by IHC against PKM2 or PKM1, respectively. The left column shows proximal jejunum villi in each animal while the next two columns are high magnification of the distal and proximal villus in each animal. The last column is colon epithelium. Blue arrows indicate nuclei with intense staining. Scales for each row are as indicated. (D) WT, KHK KO, and intestinal PKM2 KO mice were treated with H2O or HFCS and the intestinal epithelium was examined by western blot. (E, F) LDHa and ENO1 intensity were quantified relative to the b-Actin loading control (mice per group, left to right: 3|3|3|3|2|5). (G) Serum triglyceride (T.G.) following a lipid challenge was measured in mice fed water (H2O) or 25% HFCS via daily oral gavage for 2 weeks. Units are normalized to the initial timepoint to highlight changes in blood T.G. after the bolus (mice per group, top to bottom: 7|6|5|5). (H) After 2 weeks on diet mice were euthanized, and the gonadal fat deposits were weighed. Units represent total gonadal depot fat mass as a percentage of total body mass, normalized to H2O animals (mice per group, left to right: 14|14|10|5|4|5|5). (I) Liver was also harvested and analyzed for T.G. content per gram tissue (mice per group, left to right: 4|4|4|5|4|5|4). B, H: one-way ANOVA followed by Holm-Sidak post-test for multiple comparisons; E, F: two-way ANOVA followed by Holm-Sidak post-test for multiple comparisons; ns: not significant, *P<0.05, **P<0.01; all data represent means ± S.E.M. For gel source data, see Fig. S2 in the supplement.