Journal: Nature Communications

Article Title: FAM3D is essential for colon homeostasis and host defense against inflammation associated carcinogenesis

doi: 10.1038/s41467-020-19691-z

Figure Lengend Snippet: a Heatmaps of differentially expressed genes enriched in host defense responses. b mRNA expression levels of antibacterial peptides in the colons of WT or Fam3D −/− mice measured by real-time PCR. n = 8. c Reg3γ protein levels measured by immunofluorescent staining. n = 4 for each group. d GSEA analysis on Fam3D −/− vs WT EBSeq log 2 FC expression data of colonic epithelial samples. e Immunofluorescent staining for Fam3D and MUC2 in normal colon tissue. Data is representative of one experiment repeated three independent times. f Double staining of Acian blue and PAS. Yellow arrow: Acian blue positive materials; orange arrow: PAS single positive cells; orange star: dilated goblet cells, goblet cells enumerated in each crypt (10–20 crypts from each colon) and pixels of Alcian blue staining areas. Alcian blue staining areas and fluorescence intensity quantified by Image J based on six distal colon sections of four independent mice from each group. g Staining of high iron diamine/Alcian blue, dark brown, and black were sulfomucins, whereas blue staining indicates sialomucins, sulfomucin positive cells enumerated in each crypt (10–20 crypts from each colon) and the ratio of sialomucins to sulfomucins is calculated. Data is presented as the mean ± SEM. Scale bar = 50 μm. h Representative Alcian blue staining of Carnoy’s-fixed colonic sections. The thickness of the inner mucus layer (light blue band between luminal content and the mucosa) was quantified (5–10 fields from each colon) by Image J. n = 8. i Representative dual staining for UEA-I (immunofluorescence, green) and bacteria (fluorescence in situ hybridization, red), the latter using the universal EUB338 probe, on Carnoy’s-fixed colonic sections. n = 8. j PAS/Alcian blue staining of the colons from WT and Fam3D −/− mice with different ages. n = 4. Data is representative of one experiment repeated three independent times. k Quantitation of crypt length of WT and Fam3D −/− mice at different ages. Data is presented as the mean ± SEM. Statistical significance was determined by unpaired, two-tailed Student’s t test.

Article Snippet: Rat anti-EpCAM antibody (sc-53532), mouse anti-β-catenin antibody (sc-7963), rabbit anti-mouse Mucin 2 (MUC2) antibody (H300) were purchased from Santa Cruz (Dallas, TX).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Staining, Double Staining, Fluorescence, Immunofluorescence, Bacteria, In Situ Hybridization, Quantitation Assay, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: The therapeutic effect of lily polysaccharide on colitis. (a) Changes of mice weight. (Day x/Day 0 *100%, n = 5). (b) Disease activity index. ( n = 4). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1 β , IL-6 and TNF- α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: Fecal bacteria transplantation experiments showed that LP could positively regulate the gut microbiota. (a) Changes of mice weight. (Day x/Day 0 *100%, n = 5). (b) Disease activity index ( n = 5). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Bacteria, Transplantation Assay, Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: Replenishing N8AS can effectively alleviate enteritis. (a) Changes of mice weight.(Day x/Day 0 *100%, n = 5). (b) Disease activity index. ( n = 5). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. (k–o) The protein expression levels of cGAS, STING, p-TBK, p-IRF3 were assessed by western blot analysis. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Western Blot

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: In vitro verification, lily polysaccharide can inhibit the cGAS-STING pathway and play an anti-inflammatory role. (a) IL-1β mRNA expression. (b) TNF-α mRNA expression level. (c) IL-6 mRNA expression level. (d) ZO-1 mRNA expression level. (e) occludin mRNA expression level. (f) MUC2 mRNA expression level. (g–k) The protein expression levels of cGAS, STING, p-TBK, p-IRF3 were assessed by western blot analysis. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: In Vitro, Expressing, Western Blot

Journal: Acta biomaterialia

Article Title: Use of hydrogel scaffolds to develop an in vitro 3D culture model of human intestinal epithelium.

doi: 10.1016/j.actbio.2017.08.035

Figure Lengend Snippet: Fig. 7. Immunohistochemistry staining (brown) of MUC2 and MUC5AC; A: monolayer and IgG as a negative control. HT29-MTX cells layered on or suspended within B: alginate, C: L-pNIPAM, and D: L-pNIPAM-co-DMAc hydrogels under static or dynamic culture conditions at a cell density of 2 106 cells/ml following 21 days. Cell nuclei were stained with haematoxylin (blue). Yellow arrows indicate positively stained cells. Scale bar = 100 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Immunohistochemistrywas performed to investigate: brush border differentiation using CD10 antibody (1:100 rabbit polyclonal, enzyme antigen retrieval) (Abcam, Cambridge, UK); Zonulin 1 (ZO-1) protein expression which is a tight junction protein expressed by enterocytes using ZO-1 antibody (1:50, enzyme antigen retrieval) (Abcam, Cambridge, UK); enterocyte differentiation markers: alkaline phosphatase (ALP) antibody (1:200 rabbit polyclonal, heat antigen retrieval) (Abcam, Cambridge, UK), dipeptidyl peptidase IV (DPP IV) antibody (1:50mousemonoclonal, enzyme antigen retrieval) (Abcam, Cambridge, UK); and sucraseisomaltase antibody (SI) (1:50, mouse monoclonal antibody, heat antigen retrieval) (Santa Cruz, Heidelberg, Germany); HT29-MTX differentiation was assessed using MUC2 antibody (1:100 rabbit polyclonal, heat antigen retrieval) (Santa Cruz, Heidelberg, Germany) and MUC5AC antibody (1:200, mouse monoclonal antibody, heat antigen retrieval) (Abcam, Cambridge, UK).

Techniques: Immunohistochemistry, Staining, Negative Control

Journal: Mucosal immunology

Article Title: Hypoxia-inducible factor 1 in dendritic cells is crucial for the activation of protective regulatory T cells in murine colitis.

doi: 10.1038/mi.2015.67

Figure Lengend Snippet: Figure 6 Dendritic cell-specific knockout of hypoxia-inducible factor-1a (HIF-1a) stimulates mucosal epithelium to increased production of mucins in dextran sodium sulfate (DSS) colitis. Colonic mRNA expression of (a) MUC1, (b) MUC2, and (c) MUC3 was significantly higher in DSS-treated CD11cCre/HIF-1a þ f/ þ f mice than in HIF-1a þ f/ þ f DSS-treated mice or in mice not treated with DSS. (d) Sections (4 mm) of colon tissue were stained with MUC2 (red) and 4’,6-diamidino-2-phenylindole (DAPI; blue). After treatment with DSS, the amount of colonic MUC2 protein was higher in CD11cCre/HIF- 1a þ f/ þ f mice than in HIF-1a þ f/ þ f mice (original magnification 200; scale bar ¼ 50 mm). N ¼ 5–7 per group. *Po0.05; **Po0.01; ****Po0.0001. C, control; NS, not significant.

Article Snippet: MUC2 was detected with a rabbit anti-mouse MUC2 antibody (Santa Cruz Biotechnology).

Techniques: Knock-Out, Expressing, Staining, Control

Journal: bioRxiv

Article Title: Phosphatase-independent suppression of mucosal inflammation and disease progression by Salmonella SopB

doi: 10.1101/2024.09.02.610793

Figure Lengend Snippet: (A) Small intestinal tissue sections stained with H&E of non-infected (n=3) neonates or neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. Bar=100 µm. (B) Quantification of the depth of the lamina propria as a measure of the tissue edema measured in 15-20 different areas of one tissue section per animal (non-infected neonates (n=3) or neonates infected with wt (n=3) or Δ sopB (n=3)). (C) Muc2 immunostaining (red) in small intestine tissue sections of non-infected (n=3) neonates and the neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. Counterstaining with WGA (white), DAPI (blue); autofluorescence (green). Bar=50 µm. (D) Percentage of goblet cells among intestinal epithelial cells in non-infected (n=3) neonates and neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. 10-20 images with the size of 312µm x 250µm were evaluated on one tissue section per animal. (E) Goblet cell size in tissue sections of uninfected (n=3) neonates and neonates infected with wt (n=3) or Δ sopB (n=3) S . Typhimurium at day 2 p.i.. 11-21 goblet cells (µm 2 ) were analyzed on each section. (F) TUNEL staining (red) of neonatal small intestine tissue sections of non-infected (n=2) neonates and neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i.. White boxes indicate enlarged images (i, ii). Counterstaining with DAPI (blue). Bar=50 µm; insert, 20 µm. (G) Number of TUNEL positive cells in tissue sections of uninfected (n=2) neonates and the neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i. 9 -12 images with the size of 624µm x 501µm were evaluated per animal. (H) Ki67 (red) immunostaining on small intestine tissue sections of the neonates infected with wt (n=3) or Δ sopB (n=4) S . Typhimurium at day 2 p.i.. Counterstaining with E-cadherin (green), WGA (white), and DAPI (blue). Bar=50 µm, insert: 20 µm. (I) Number of Ki67 positive cells in small intestinal tissue sections of uninfected (n=2) neonates and the neonates infected with wt (n=5) or Δ sopB (n=4) S . Typhimurium at day 2 p.i. 5-29 crypts were analyzed per section. Representative images are shown. Quantified data are shown as individual points in violin plots, solid lines represent the median. Data were analyzed using One-way ANOVA Kruskal–Wallis test with Dunn’s multiple comparisons post-test. ns, non-significant; *, p<0.05; ****, p<0.0001.

Article Snippet: Rabbit anti-Ki67 (Ab15580, Abcam), mouse anti-E-cadherin (610182, BD Transduction Laboratories), rat anti-PMN (Ly6-6B2, SeroTec), rabbit anti-Muc2 (GTX100664, BIOZOL) were used at the appropriate dilution followed by the fluorophore-conjugated donkey secondary antibody (Jackson ImmunoResearch).

Techniques: Staining, Infection, Immunostaining, TUNEL Assay

Journal: Evidence-based complementary and alternative medicine : eCAM

Article Title: Liangxue Tongyu Prescription Alleviates Brain Damage in Acute Intracerebral Hemorrhage Rats by Regulating Intestinal Mucosal Barrier Function.

doi: 10.1155/2022/2197763

Figure Lengend Snippet: Figure 6: Efect of LTP on MUC2, SIgA, and GPR43 expression in the colon tissue of AICH rats: (a) MUC2 expression, (b) SIgA expression, (c) GPR43 expression, and (d) WB electrophoretograms of MUC2, SIgA, and GPR43 in the colon tissue samples. ap < 0.05 vs. the blank and sham groups. bp < 0.05 vs. the model group. cp < 0.05 vs. the LTP group (n 3).

Article Snippet: Te following primary antibodies were used for western blotting: rabbit anti-ZO-1 (Termo Fisher Scientifc, 40–2200, 1 :1000), OCLN (Abcam, ab167161, 1 :1000), CLDN-5 (ABclonal, A10207, 1 :1000), CLDN-1 (Termo Fisher Scientifc, 71–7800, 1 :1000), SIgA (Bioss Biotech, bs-0645R, 1 :1000), MUC2 (Boster Biotech, BM5029, 1 :1000), GPR43 (Bioss Biotech, bs-13536R, 1 :1000), and a βactin antibody (ABclonal, ac026, 1 :10000).

Techniques: Expressing