Fig. 4

Long-term β-glucan supplementation prevented colonic mucosa barrier impairment and inflammation and ameliorated endotoxemia in HFFD-induced obese mice. a Alcian blue-stained colonic sections were showing the mucus layer (arrows). Opposing black arrows with shafts delineate the mucus layer that was measured (scale bar 50 μm). b The quantification of the colonic mucus layer was statistically analyzed (per section/2 sections per animal, n = 5). c Immunofluorescence images of colonic sections stained with Anti-MUC2 antibody and DAPI. Opposing white arrows with shafts delineate the mucus layer. Inset (HFFD group) shows a higher magnification of bacteria-sized, DAPI-stained particles in closer proximity to host epithelium and even crossing this barrier. Scale bar 50 μm, inset 10 μm. d FISH analysis of sections of the colon using the general bacterial probe EUB338-Alexa Fluor 488 (green), and nuclear staining DAPI (blue). Arrows indicate the distance between bacteria and epithelium. Scale bar 20 μm. e Quantitation of colonic Reg3γ by RT-PCR (n = 6). f Protein levels of occludin and ZO-1 in the colon (n = 5). g Serum endotoxin level (n = 10). h mRNA expression of TNF-α, IL-1β, and IL-6 in the colon (n = 5). i The quantification of colon length was statistically analyzed (n = 10) and representative images of colons. j TNF-α, IL-6, and IL-1β levels in the serum (n = 10). Values are mean ± SEM. ^*p < 0.05 vs. control (Con). ^#p < 0.05 vs. high-fat and fiber-deficient diet (HFFD)