Extended Data Fig. 5. Proteomics and metabolomics analyses reveal downregulation of lipid biosynthesis and chylomicron production, increased lipid droplet formation and suppression of mitochondrial metabolism in DARS2-deficient enterocytes.
a, Volcano plot presenting the proteome landscape comparing DARS2tamIEC-KO(n = 11) to Dars2fl/fl (n = 9) mice 7 days after the last tamoxifen injection. b, Immunoblot analysis with the indicated antibodies of protein extracts from proximal SI IECs isolated from Dars2fl/fl and DARS2tamIEC-KO mice 7 days after the last tamoxifen injection. α-tubulin and vinculin were used as loading controls. c-d, Emapplots of the Over Representation Analysis (ORA) performed on the significantly upregulated (Log2FC > 0.5 and p.adj < 0.05) proteins (c) and on the significantly downregulated (Log2FC < −0.5 and p.adj<0.05) proteins (d) when comparing DARS2tamIEC-KO (n = 11) to Dars2fl/fl (n = 9) mice 7 days upon tamoxifen injection. e, Volcano plot illustrating the differential intracellular metabolite levels comparing SI IECs of DARS2tamIEC-KO (n = 15) to Dars2fl/fl (n = 9) mice 7 days upon the last tamoxifen injection. f, Each data point in the bar graph represents the mean ± s.e.m of three technical replicates of one animal and is expressed as AUs relative to the average value of all control mouse samples for each metabolite detected in IECs from DARS2tamIEC-KO (n = 15) to Dars2fl/fl (n = 8) mice 7 days upon the last tamoxifen injection. P values were calculated by unpaired two-sided Welch’s Student t-test with S0 = 0.1 and a permutation-based FDR of 0.01 with 500 randomizations (a), one-sided Fisher’s exact test with Benjamini-Hochberg multiple-testing correction (c, d), one-sided Student t-test with Benjamini-Hochberg multiple-testing correction (e) and unpaired two-sided Student’s t-test with no assumption of equal variance or two-sided nonparametric Mann-Whitney U -test (f).