Fig. 4: HFHSD induces hyperproliferation of ISCs and progenitors.
From: Diet-induced alteration of intestinal stem cell function underlies obesity and prediabetes in mice
a, Distribution of cycling cells across CD- and HFHSD-derived cell clusters depicted as cell densities projected onto the UMAP plot and quantified as proportions of cells in each cell-cycle stage. Cells were classified using a cell-cycle score, calculated using the expression of genes related to cell cycle. Data are mean ± s.e.m. of biologically independent samples, n = 3 mice per group. b, Distribution of ISCs and progenitors over the three cell-cycle stages visualized as cell densities in a scatter-plot of S- versus G2/M-phase score levels. Higher score levels indicate higher expression of involved genes. Dotted lines depict classification borders. Densities are Gaussian kernel estimates. c, Proportions of cycling cells in CD and HFHSD-derived ISCs and progenitor clusters. Table indicates percentages. Data are shown as mean ± s.e.m. of biologically independent samples, n = 3 mice per group. EP, enterocyte progenitor; GP, goblet progenitor. d, Heatmap of mean expression values per cluster of selected genes used for cell-cycle scoring (black) or highly correlating with S and G2M scores (grey, Pearson correlation >0.7). Only cells classified as cycling (S or G2M phase) are shown. * Indicates differentially expressed genes between CD and HFHSD conditions (two-sided, limma, adjusted P < 0.01, logFC > 0.1), n = 3 mice per group. FC, fold change. P values are provided in Supplementary Table 5. e,f, Representative LSM images (e) and quantification (f) of EdU incorporation after a 2-h EdU (white) pulse in the TA zone in duodenal sections of CD- and HFHSD FVF mice. Lyz1+ Paneth cells are shown in red. Scale bar, 25 µm, n = 4 mice per group. Data are mean ± s.e.m. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test. FDR, false discovery rate. g, Determination of Lgr5–EGFPhi cells from CD- and HFHSD-fed Lgr5-ki mice by flow cytometry, n = 4 mice per group. Data are mean ± s.e.m. of biologically independent samples. h,i, Representative LSM images from cytospin (h) and quantification (i) of EdU+Lgr5–EGFPhi cells from CD- and HFHSD-fed Lgr5-ki mice after a 2-h EdU (white) pulse. Scale bar, 40 µm, n = 4 CD mice, n = 3 HFHSD mice. Data are mean ± s.d. of biologically independent samples. Statistical significance was determined by two-tailed Student’s t-test.
