Extended Data Fig. 2: Enhanced glucose consumption in Kdm2a KO mice under cold insult.
From: Kdm2a inhibition in skeletal muscle improves metabolic flexibility in obesity
(a)The food and water intake during metabolic cages monitoring of cold-adapted Kdm2a fl/fl and Kdm2aSKO mice (n = 8 per group). (b) Muscle mass analysis between ND Kdm2a fl/fl and Kdm2aSKO mice with cold challenge (n = 8 per group). (c) Representative H&E staining images in slices of cold-adapted Kdm2a fl/fl and Kdm2aSKO mice Gas tissues, average myotubes area analysis, Arrows indicated the blood vessels, Scale bars, 50 μm, (n = 5 per group). (d) Representative immunostaining of Laminin (green), CD31 (red) and DAPI (blue) in Gas tissue cross sections of cold-adapted Kdm2a fl/fl and Kdm2aSKO male mice, CD31+ number analysis (right), Scale bars, 50 μm, (n = 5 per group). (e) Representative immunostaining of Laminin (red), MHC II (green) and DAPI (blue) in Gas tissue cross sections of cold-adapted Kdm2a fl/fl and Kdm2aSKO male mice. Scale bars, 50 μm, (n = 4 per group). (f) Body weight gain of Kdm2a fl/fl and Kdm2aSKO mice fed with ND without cold challenge (n = 5 per group). (g) IGTT and (h) ITT of Kdm2a fl/fl and Kdm2aSKO mice fed with ND without cold challenge (n = 4/5 per group). AUC analysis (right). (i) Western blot analyses Sdhb, Ndufs1, Ndufs3 and Pgc1a of ND Kdm2a fl/fl and Kdm2aSKO mice without cold challenge (n = 4 per group). Protein quantitative analysis (j). (k) Western blot analysis of H3K36me2 and Kdm2a in Kdm2a-WT and Kdm2a-KO C2C12 myotubes (n = 4 per group). Protein quantitative analysis (right). (l-o) Treat Kdm2a-WT and Kdm2a-KO C2C12 myotubes with 2 hours of cold stimulation. (l) Glucose uptake assay in Kdm2a-WT and Kdm2a-KO C2C12 myotubes with cold stimulation (n = 10). (m) RT-qPCR analysis of Mb, Vegfa and Myh7 expression (n = 6 per group). (n) Western blot analysis of Myh4, Sdhb, Ndufs1, Ndufs3, Pgc1a (n = 4 per group). Protein quantitative analysis (right). (o) mtDNA copy number analysis (n = 6 per group). Data were expressed as mean ± SEM. Statistical significance was assessed by two-way ANOVA (f-h) followed with Bonferroni’s multiple comparisons test or two-sided Student’s t test (a-e, i-o).
