Spatial transcriptomic analyses of distal colon cross-sections from mice fed high fiber and control diet

To perform a global analysis of high fiber diet-induced changes in gene expression in various compartments of the colonic microenvironment, we employed a spatial transcriptomic analysis using the 10X Genomics® platform. This technique allowed for visualization of gene expression in histological cross-sections of the distal colon with spatial information and therefore enabled detection of differentially expressed genes simultaneously in diverse cell types, together with information about their anatomical location in the tissue. Distal colon samples were processed using a Visium Spatial Gene Expression Slide & Reagent Kit (PN-1000184, 10X Genomics) as per the manufacturer's instructions. Unbiased clustering divided the colon into four distinct zones: epithelium, lamina propria & submucosa, muscularis, and neuronal plexus, as visualized by hematoxylin and eosin (H&E) staining. Overall design: Four samples from two control diet-fed mice and four samples from two high fiber diet-fed mice were used