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: Frontiers in Physiology

Article Title: Characterization of Mucus-Related Properties of Streptococcus thermophilus: From Adhesion to Induction

doi: 10.3389/fphys.2018.00980

Figure Lengend Snippet: Characterization of the colonic epithelial response in S. thermophilus LMD-9 mono-colonized rats. Germfree (GF) rats were inoculated with sterile milk (GF, n = 5) or with 5 × 10 8 CFU of a culture of S. thermophilus LMD-9 (Ino-LMD9, n = 6) and euthanized 30 days after inoculation. Similar groups GF +lac ( n = 6) and Ino-LMD9 +lac ( n = 6) were given tap water containing 4.5% (w/v) lactose (A) Experimental design: establishment of S. thermophilus LMD-9 (with or without lactose) in the gastrointestinal tract was monitored by enumeration of the bacterial counts in the feces. The lactate concentration was measured in the cecum. (B) Representative western blot and densitometric analyses of KLF4 protein in GF and Ino-LMD9 samples. (C) Representative western blot and densitometric analyses of KLF4 protein in GF +lac and Ino-LMD9 +lac samples. (D) Representative images, counts of goblet cells stained with Alcian Blue (indicated as AB) and counts of total number of cells per crypt stained with Hematoxylin from in GF +lac and Ino-LMD9 +lac samples; scale bar: 50 μm. (E) Representative images, Muc2 immunostaining in GF +lac and Ino-LMD9 +lac rats; scale bar: 50 μm. (F) muc2 mRNA expression level in GF +lac , Ino-LMD9 and Ino-LMD9 +lac rats, and counts of the number of muc2 positive cells per crypt GF +lac and Ino-LMD9 +lac rats. The asterisk indicates a statistical difference relative to GF rats ( p < 0.05); ns: not significant.

Article Snippet: Muc2 mRNA was analyzed using TaqMan Gene Expression Assays (Rn01498197_m1, Applied Biosystems.

Techniques: Sterility, Concentration Assay, Western Blot, Staining, Immunostaining, Expressing

Journal: EMBO Reports

Article Title: Haploinsufficient phenotypes promote selection of PTEN and ARID1A-deficient clones in human colon

doi: 10.1038/s44319-025-00373-0

Figure Lengend Snippet: ( A ) Number of positive cells per crypt for markers used in the panel. Applied on WT crypts. Box plot shows min, median and max. ( B – U ) ANLN, S/G phase marker. pH3, mitosis marker. MUC2, goblet cells marker. Chga, enteroendocrine marker. Cleaved PARP1, apoptosis marker. ANLN, ph3 and cleaved PARP1 expressed as a number of positive cells per crypt. MUC2 and ANLN expressed as frequency out of total DAPI cells. ( Y ) Definition of crypt axis levels based on CA2 and MCM2 staining in en face embedded tissue. Different levels within a section are seen due to tissue not being completely flat during processing. CA2 + MCM2- crypt top. CA2 + MCM2+ crypt middle. CA2− MCM2+ crypt bottom. Dashed borders indicate crypt top, middle or top according to marker expression. Data information: Data presented as mean ± SD. All scale bars indicate 400 μm. A paired t -test or Wilcoxon test was performed to assess statistical significance based on the type of distribution. ( B ) p = 0.297, ( C ) p = 0.203, ( D ) p = 0.175, ( E ) p = 0.355, ( F ) p = 0.178, ( G ) p = 0.597, ( H ) p = 0.56, ( I ) p = 0.406, ( J ) p = 0.32, ( K ) p > 0.999, ( L ) p = 0.541, ( M ) p = 0.017, ( N ) p = 0.36, ( O ) p = 0.025, ( P ) p = 0.075, ( Q ) p = 0.035, ( R ) p = 0.12, ( S ) p = 0.508, ( T ) p = 0.247, ( U ) p = 0.0004, ( V ) p > 0.999. * p < 0.05, *** p < 0.001. ( B – E ) N = 23 PTEN and WT clones. ( F – I ) N = 25 ARID1A and WT clones. ( J – M ) N = 12 MAOA and WT clones. ( N – Q ) N = 14 STAG2 and WT clones. ( R – U ) N = 26 KDM6A and WT clones. ( V ) N = 10 ARID1A and WT clones.

Article Snippet: MUC2 Rabbit , Santa Cruz , SC-15334 , 1:400.

Techniques: Marker, Staining, Expressing, Clone Assay

Journal: EMBO Reports

Article Title: Haploinsufficient phenotypes promote selection of PTEN and ARID1A-deficient clones in human colon

doi: 10.1038/s44319-025-00373-0

Figure Lengend Snippet: Reagents and tools table

Article Snippet: MUC2 Rabbit , Santa Cruz , SC-15334 , 1:400.

Techniques: Modification, Amplification, Sequencing, Software, DNA Extraction, Transferring

Journal: Theranostics

Article Title: Exercise ameliorates osteopenia in mice via intestinal microbial-mediated bile acid metabolism pathway

doi: 10.7150/thno.104186

Figure Lengend Snippet: Exercise and corresponding FMT protected intestinal barrier and reduced inflammation in OVX mice. (A) Representative images showing intestinal structure by AB-PAS staining and immunofluorescence staining of Muc2 and Tjp1. Red arrows indicating goblet cells. (B) Quantitative results of goblet cells. (C) The intestinal permeability measured by circulating FITC-labeled dextran. (D-E) The mRNA expression levels of genes associate with intestinal barrier, including Muc2 and Tjp1 . (F-I) The mRNA expression levels of inflammatory factors ( Il1b , Il6 , Il17 , and Tnfa ) in small intestinal tissue. (J) Serum LPS concentration. (K-N) Serum levels of inflammatory factors, including IL1β, IL6, IL17, and TNFα. Graphs show mean ± SEM (n = 6 per group), with statistical significance determined by one-way ANOVA followed by Bonferroni's multiple comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Immunofluorescent staining of intestinal tissues was performed using Muc2 antibody (ABclonal Technology, China) and Tjp1 antibody (ABclonal Technology, China), and the cell nuclei were counterstained with DAPI.

Techniques: Staining, Immunofluorescence, Permeability, Labeling, Expressing, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Congruence of Transcription Programs in Adult Stem Cell-Derived Jejunum Organoids and Original Tissue During Long-Term Culture

doi: 10.3389/fcell.2020.00375

Figure Lengend Snippet: Organoids derived from adult intestinal stem cells show intrinsic programming to differentiate into different epithelial cell lineages and express small intestine-specific genes. (A) Cell type-specific transcripts for Crypt Base Columnar (CBC) and Stem cells, Label-retaining (LRC) +4 cells, Proliferation (Prol), Niche factors (NF), Paneth cells (PC), Goblet cells (GC), Enteroendocrine cells (EEC), and Absorptive cells or enterocytes (Abs/EC). (B) Overlaying the NCBI gene tracks of MUC2 NC_010444.4 on chromosome 2 at location 689363–719542bp (green area), shows identical overlap with the mapped reads and coverage (Cov) from organoid and tissue samples, but not in IPEC-J2 (ENS; Ensembl reference genome). To confirm MUC2 protein translation and subsequent mucus formation, Carnoy fixed tissue, organoid, and IPEC-J2 samples were stained with PAS/Alcian blue (left) and porcine anti-MUC2 (right; black and white size bars indicate 100 μm). (C) Most small intestine-specific genes identified in the human protein atlas are also expressed in porcine jejunum tissue and their derived organoids (>74%), whereas fewer are expressed in ileum organoids (52%). IPEC-J2 only expressed 32% of the small intestine-specific genes. (D) Organoid transcriptomes from jejunum and ileum were compared to identify differences in expressed genes, showing large overlap of genes (Venn), but also some differences. The different genes were analyzed using TOPPfun to identify putative differences in gene ontology and pathways (TMT, Transmembrane transport; PSP, Peptidyl-serine phosphorylation).

Article Snippet: After cutting 5 μM-thick sections and subsequent drying on glass slides, sections were rehydrated, blocked in 5% normal goat serum, and stained using anti-MUC2 antibody (1:200, AB_1950958, GeneTex) overnight.

Techniques: Derivative Assay, Staining, Phospho-proteomics