Fig. 5: Fatty acid synthesis and Ppar signalling are upregulated on HFHSD.
From: Diet-induced alteration of intestinal stem cell function underlies obesity and prediabetes in mice
a, Heatmap-based clustering analysis of the 297 discriminative metabolite masses (FC ≥ 2 and P ≤ 0.05). Each coloured cell on the map corresponds to an intensity value, with samples in rows and features in columns. Euclidean distance and Ward’s method were applied for clustering analysis. Statistical significance was determined by two-tailed Student’s t-test. b, Pathway enrichment analysis of deregulated metabolites was performed with MetaboAnalyst 3.0. The enrichment method was hypergeometric test. Topology analysis was based on relative betweenness centrality. The P value was calculated from the enrichment analysis without adjustment (FC ≥ 2 and P ≤ 0.05). Metabolic pathways are represented as circles according to their scores from enrichment (vertical axis) and topology analyses (pathway impact, horizontal axis). c, Overview of experimental design for data integration. d, Ingenuity pathway analysis showing overlap of significantly deregulated metabolites on HFHSD from MALDI–MSI profiling and genes from microarray analysis. Shown are values from microarray analysis and the red line indicates the significance cutoff. e, Enriched KEGG pathways in genes differentially regulated between CD and HFHSD conditions (Enrichr, Fisher’s exact test, two-sided). Genes with FDR < 0.01 and logFC > 0.1 were considered and weighted by logFC. f, Volcano plots showing differential expression and its significance (−log10(FDR), limma) for each gene on HFHSD compared to CD. Red lines indicate thresholds used for significance level and gene expression change and regulated genes are highlighted in black. Annotated genes are the top ten genes ranked by FDR. g, Mean expression levels for selected genes. * Indicates a significant change (limma, FDR < 0.01, logFC > 0.1). h,i, Protein expression analysis by western blot in cytoplasmic (cyto) and nuclear (nucl) extracts from SI crypts of CD- and HFHSD-fed FVF mice. Representative immunoblots (h) and relative quantification of band signal intensity (i), n = 4 mice per group (Srebp1, Scd1, Acc, Fasn) and n = 7 mice per group (Pparγ). Data are presented as box-and-whisker plots. The lower and upper boundaries of the boxes represent the 25th and 75th percentiles, respectively. The centre lines indicate the medians and whiskers represent the maximum and minimum values. Statistical significance was determined by two-tailed Student’s t-test. Circles represent biological independent samples. norm., normalized.
