primary antibody against muc2  (Proteintech)

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Intestinal Muc2 knockout aggravated pancreatic injuries in AP. A PAS-staining and the numbers of mucus-containing goblet cells in the duodenum of AP patients with different etiologies. Original magnification, ×200. B Immunofluorescence and quantification of MUC2 in the duodenum of AP patients with different etiologies. Original magnification, ×200. C Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). D Serum amylase activity. E TUNEL staining and quantification of TUNEL + cells in pancreas. Original magnification, ×200. F Real-time PCR analysis of IL-1β, TNF-α, IL-6 mRNA levels in pancreas; tubulin was used as the reference gene. G ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. The data are presented as the means ± SEM ( n = 6 mice per group). ns, no significant difference; * p < 0.05; HC, healthy controls; ABP, acute biliary pancreatitis; HTGAP, hypertriglyceridemic acute pancreatitis; AP, acute pancreatitis (cerulein); WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, Staining, Immunofluorescence, Activity Assay, TUNEL Assay, Real-time Polymerase Chain Reaction, Analysis, Enzyme-linked Immunosorbent Assay, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Intestinal Muc2 knockout aggravated AP-related intestinal injuries. A Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). B TUNEL staining and quantification of TUNEL + cell in the colon. Original magnification, ×200. C Immunofluorescence staining for Claudin-1 and Occludin (green) in the colon. Original magnification, ×100. D Real-time PCR analysis of IL-1β, TNF-α, IL-6 mRNA levels in the colon; tubulin was used as the reference gene. E ELISA analysis of serum DAO, D-lactate levels. F FISH detection of bacteria translocation in the pancreas and colon by the EUB338 probe. Original magnification, ×200. G Colony-forming units (CFUs) of anaerobic bacteria in mesenteric lymph nodes and liver. The data are presented as the means ± SD ( n = 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelial cells; AP, acute pancreatitis. FISH, fluorescence in situ hybridization

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, TUNEL Assay, Staining, Immunofluorescence, Real-time Polymerase Chain Reaction, Analysis, Enzyme-linked Immunosorbent Assay, Bacteria, Translocation Assay, Control, Fluorescence, In Situ Hybridization

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Muc2 deficiency aggravated intestinal microbiota dysbiosis and reduced riboflavin production. A The bar graph of the relative abundance of the top 20 genera in feces between WT and Muc2 ΔIEC groups. B The bar graph of the relative abundance of the top 20 genera in feces between AP and Muc2 ΔIEC + AP groups. C KEGG enrichment analysis of differential fecal metabolites between WT and Muc2 ΔIEC groups. D KEGG enrichment analysis of differential fecal metabolites between AP and Muc2 ΔIEC + AP groups. E The VIP value plots of the differential fecal metabolites (the top 20 of VIP value) between WT and Muc2 ΔIEC groups. F The VIP value plots of the differential fecal metabolites (the top 20 of VIP value) between AP and Muc2 ΔIEC + AP groups. G Riboflavin, flavin mononucleotide, and flavin adenine dinucleotide levels in feces. H Targeted metabolomics analysis of riboflavin, flavin mononucleotide and flavin adenine dinucleotide levels in pancreas. I Network analysis between gut microbiome and fecal metabolites. Data are presented as the means ± SEM ( n = 5 or 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Analysis, Microbiome, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Rib deficiency weakened the protective effect of Lactobacillus plantarum on AP in Muc2 ΔIEC mice. A Schematic diagram of L. plantarum AR113 with Rib deletion (AR113Δ Rib ). B Experiment design and time axis of AR113Δ Rib gavage and AP induction in Muc2 ΔIEC mice. C Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). D Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). E ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. F Real-time PCR analysis of IL-1β, TNF-α, and IL-6 mRNA levels in the colon. Tubulin was used as the reference gene. G Immunofluorescence staining for claudin1and occludin (green) in the colon. Original magnification, ×200. H FISH detection of bacteria translocation in the pancreas by the EUB338 probe. Original magnification, ×200. The data are presented as the means ± SEM ( n = 6 mice per group). * p < 0.05; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AR113, L. plantarum AR113; AR113Δ Rib , L. plantarum AR113 with Rib deletion; AP, acute pancreatitis; FISH, fluorescence in situ hybridization

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Enzyme-linked Immunosorbent Assay, Analysis, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Bacteria, Translocation Assay, Fluorescence, In Situ Hybridization

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Riboflavin alleviated AP in Muc2 ΔIEC mice. A Schematic diagram of riboflavin treatment. B Histopathological changes and pathological scores of the pancreas. Original magnification, ×100 (top panels) or ×200 (bottom panels). C Serum amylase activity. D ELISA analysis of serum IL-1β, TNF-α, and IL-6 levels. E Histopathological changes and pathological scores of the colon. Original magnification, ×100 (top panels) or ×200 (bottom panels). F Real-time PCR analysis of IL-1β, TNF-α, and IL-6 mRNA levels in the colon. Tubulin was used as the reference gene. G Immunofluorescence staining for claudin-1and occludin (green) in the colon. Original magnification, ×200. H FISH detection of bacteria translocation in the pancreas and colon by the EUB338. I CFUs of anaerobic bacteria in liver and mesenteric lymph nodes. The data are presented as the means ± SEM ( n = 6 mice per group). * p < 0.05; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis; RF, riboflavin; CFUs, colony-forming units

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Analysis, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Bacteria, Translocation Assay

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Riboflavin reduced the pro-inflammatory macrophage infiltration in AP mice with intestinal Muc2 knockout. A Immunofluorescence staining and quantification for pro-inflammatory macrophages (F4/80 + NOS2 + ) in the pancreas. Original magnification, ×200. B Immunofluorescence staining and quantification for neutrophil (Ly6G + ). Original magnification, ×200. C Flow cytometry analysis and quantification of pro-inflammatory macrophages (F4/80 + CD86 + ) and anti-inflammatory macrophages (F4/80 + CD206 + ) in the pancreas. D Flow cytometry analysis and quantification of neutrophil (CD11b + Ly6G +) . E Immunofluorescence staining and quantification for Th17 cells (CD4 + RORγ + ). Original magnification, ×200. F Immunofluorescence staining and quantification for Treg cells (CD4 + FOXP3 + ). Original magnification, ×200. G Immunofluorescence staining and quantification for pro-inflammatory macrophages (F4/80 + NOS2 + ) in the pancreas. Original magnification, ×200. H Real-time PCR analysis of Nos2 mRNA levels in macrophage with different concentrations of riboflavin. Tubulin was used as the reference gene. I Real-time PCR analysis of Nos2, TNF-α, IL-6, and IL-1β mRNA levels. Tubulin was used as the reference gene. J Immunofluorescence staining for NOS2 (red). Original magnification, ×200. The data are presented as the means ± SEM ( n = 6 mice per group). ns, no significant difference; * p < 0.05; WT, Muc2 flox/flox littermate control; Muc2 ΔIEC , mice with specific deletion of Muc2 in intestinal epithelium; AP, acute pancreatitis; RF, riboflavin

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Knock-Out, Immunofluorescence, Staining, Flow Cytometry, Analysis, Real-time Polymerase Chain Reaction, Control

Journal: Microbiome

Article Title: Gut microbiota-derived riboflavin mediates protective effects of MUC2 in acute pancreatitis via CD40-macrophage pathway

doi: 10.1186/s40168-026-02375-z

Figure Lengend Snippet: Reduced intestinal MUC2 was associated with decreased serum riboflavin in AP patients. A Spearman correlation analysis of intestinal MUC2 expression level and serum riboflavin concentration in the initial patient cohort. B Analysis of intestinal MUC2 expression levels between MAP/MSAP and SAP. C Analysis of serum riboflavin concentrations between MAP/MSAP and SAP. D Spearman correlation analysis of intestinal MUC2 expression level and Ranson score in the initial patient cohort. E Spearman correlation analysis of serum riboflavin concentration and Ranson score in the initial patient cohort. F Spearman correlation analysis of serum riboflavin concentration and Ranson score in the validation cohort. AP, acute pancreatitis; MAP, mild acute pancreatitis; MSAP, moderately severe acute pancreatitis; SAP, severe acute pancreatitis

Article Snippet: Membranes were blocked with 3% bovine serum albumin (BSA) for 1 h, then incubated overnight at 4 °C with primary antibodies against MUC2 (A14659; Abclonal, China) or CD40 (A22714; Abclonal, China) in primary antibody dilution buffer (Epizyme Biotech, China).

Techniques: Analysis, Expressing, Concentration Assay, Biomarker Discovery

Journal: BMC Veterinary Research

Article Title: Dietary capsaicin supplementation attenuates jejunal injury of piglets challenged with lipopolysaccharide

doi: 10.1186/s12917-026-05346-6

Figure Lengend Snippet: Effect of dietary CAP supplementation on jejunal barrier function of LPS-challenged piglets. A The mRNA expression of jejunal ZO-1 and OCLN ( n = 8). B , C Representative immunofluorescence results and quantitative analysis of MUC2 protein in the jejunum (scale bar = 100 μm; n = 5). CON, piglets fed a basal diet; LPS, LPS-challenged piglets fed a basal diet; LCA, LPS-challenged piglets fed a basal diet with 800 mg/kg CAP. a, b Different letters among groups were considered as statistical significance ( P < 0.05). The results were emerged as mean and SEM. ZO-1 Zonula occludens-1, OCLN Occluding, MUC2 Mucin 2

Article Snippet: Subsequently, the slides were blocked with 3% bovine serum albumin for 30 min and then incubated with the primary antibody against MUC2 (#GB11344-100; Servicebio Technology) in a wet box overnight at 4 °C.

Techniques: Expressing, Immunofluorescence

Journal: Gut Microbes

Article Title: Microbiota shape the colon epithelium controlling inter-crypt absorptive goblet cells via butyrate–GP R109A signalling

doi: 10.1080/19490976.2025.2573045

Figure Lengend Snippet: Absorptive inter-crypt goblet cells are intricately linked to the microbiota. (A) Schematic representation of the experimental design illustrating the AIMD mix treatment performed in SPF mice. (B) Immunofluorescence staining of proximal colonic sections from control and AIMD mice using anti-CA1 and UEA1 lectin for mucus and DAPI for DNA counterstaining. Quantification of CA1 + UEA1 + cells within total CA1 + cells. Scale bar, 50 µm; n = 5 per group. (C) Whole-mount proximal colon from control and AIMD mice stained with the UEA1 lectin for mucus and CellMask for the cell membrane. Representative Z-stacks from the region outlined by the white box are shown at the bottom. UEA1 + cells in inter-crypt region are indicated with arrows. Scale bars, 20 µm. (D) Staining of proximal colonic sections from control and AIMD mice with the UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in the inter-crypt region are indicated with white arrows and the epithelial surface is indicated by the white dashed line. Quantitation of these cells is shown on the right. Scale bar, 50 µm; n = 5 in each group. (E) Volcano plot showing DEGs in the absorptive populations from GF and SPF mice. Some goblet cell markers are highlighted in the figure. The Wilcoxon statistical test with a significance threshold of adjusted p < 0.05 and log2-fold change > 0.25 was used. (F) UMAP plot of the scRNA-seq data showing the Muc2 + population (red dots) within the absorptive cluster in both the GF and SPF mice, grey dots represent canonical absorptive cells (enterocytes and immature enterocytes). The right panel shows the proportion of bona fide absorptive inter-crypt goblet cells. (G) Schematic representation of the experiment with FMT from SPF to GF mice. (H) Immunofluorescence staining of proximal colonic sections from SPF colonized and GF mice with anti-CA1, UEA1 lectin for mucus, and DAPI for DNA counterstaining. UEA1 + cells in the intercrypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Quantification of CA1 + UEA1 + cells within total CA1 + cells. Scale bar, 50 µm; n = 8 per group. (I) Staining of proximal colonic sections from SPF colonized and GF mice with UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by the white dashed line. Quantitation of UEA1 + cells per crypt is shown on the right. Scale bar, 50 µm; n = 8 in each group. (J) Schematic representation of the experimental design illustrating the ABx (antibiotics cocktail) treatment performed in GF mice. (K) Proximal colonic sections from GF control and GF treated with antibiotics-cocktail mice stained with UEA1 lectin for mucus and DAPI for DNA. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by the white dashed line. Quantitation of UEA1 + cells per crypt is shown on the right. Scale bar, 50 µm; n = 4 in each group. All error bars are SEM.

Article Snippet: Sections from formalin-fixed paraffin-embedded (FFPE) samples were used for immunofluorescence with primary antibodies against MUC2 (Santa Cruz Biotechnology #sc-7314, 1:50) and CA1 (Santa Cruz Biotechnology #sc-393490, 1:200).

Techniques: Immunofluorescence, Staining, Control, Membrane, Quantitation Assay

Journal: Gut Microbes

Article Title: Microbiota shape the colon epithelium controlling inter-crypt absorptive goblet cells via butyrate–GP R109A signalling

doi: 10.1080/19490976.2025.2573045

Figure Lengend Snippet: Microbiota-derived butyrate reduces absorptive inter-crypt goblet cell numbers through GPR109A. (A) Schematic representation of the experiment using the inulin-rich and control diet treatments performed in SPF mice. (B) Volcano plots highlighting DEGs within the absorptive cell populations of inulin-rich and control diet-fed mice. Some goblet cell markers are highlighted in the figure. The Wilcoxon statistical test with a significance threshold of adjusted p < 0.05 and log2-fold change > 0.25 was used. (C) UMAP plot of the scRNA-seq data highlighting the Muc2 + cell population within the absorptive cluster in both inulin-rich and control diet-fed mice. The gray dots represent canonical absorptive cells (enterocytes and immature enterocytes), while red dots represent Muc2 + cells. The right panel shows the proportion of Muc2 + cells. (D) Staining of proximal colon sections from inulin-rich and control diet-fed mice with the UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar 50 µm, n = 4 in each group. (E) Schematic representation of the experimental design illustrating the inulin-rich diet administration and mono-colonization experiments performed in GF mice. (F) Staining of proximal colon sections from B. ovatus or B. thetaiotaomicron ('B. theta') mono-colonized mice fed an inulin-rich diet with UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar, 50 µm; n = 5 in each group. (G) Schematic representation of the experiment using tributyrin supplementation of GF mice. (H) Staining of proximal colon sections from vehicle or tributyrin supplemented GF mice with UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar 50 µm; n = 4/5 in each group. (I) Staining of proximal colon sections from Gpr109a WT and KO mice with the UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar 50 µm, n = 8/6 in each group. All error bars are SEM.

Article Snippet: Sections from formalin-fixed paraffin-embedded (FFPE) samples were used for immunofluorescence with primary antibodies against MUC2 (Santa Cruz Biotechnology #sc-7314, 1:50) and CA1 (Santa Cruz Biotechnology #sc-393490, 1:200).

Techniques: Derivative Assay, Control, Staining

Journal: Gut Microbes

Article Title: Microbiota shape the colon epithelium controlling inter-crypt absorptive goblet cells via butyrate–GP R109A signalling

doi: 10.1080/19490976.2025.2573045

Figure Lengend Snippet: Absorptive inter-crypt goblet cell numbers are associated with aging (A) Staining of proximal colon sections from aged (~80 weeks old) and young (8–12 weeks old) mice with UEA1 lectin for mucus and DAPI for DNA counterstaining. UEA1 + cells in the inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar 50 µm, n = 8/5 in each group. (B) Proportion of Muc2 + cells in the absorptive cluster in aged (>65 y, n = 3) and young-adult (<65 y, n = 4) human colonic samples. Analysis was performed using the Gut Cell Atlas dataset. (C) Immunofluorescence staining of young and aged human descending colonic sections using anti-CA1 and anti-MUC2 antibodies and DAPI for DNA counterstaining. Double-stained cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line (please note that MUC2 and CA1 are expressed in different compartments within a cell). Quantification of CA1 + MUC2 + cells within total CA1 + cells on the right. Scale bar, 30 µm; n = 5/8 per group. (D) Schematic representation of the experimental design for human fecal microbiota transplantation (FMT) in germ-free (GF) mice. The donors were all healthy, female (41 and 72 y, batch 1) and male (35 and 77 y, batch 2). (E) Staining of proximal colon sections from GF mice colonized with human microbiota from aged or young donors with the UEA1 lectin for mucus and DAPI for DNA counterstaining. Two independent experiments were performed using FMT from different donors for each group, as illustrated in D. UEA1 + cells in inter-crypt region are indicated with arrows, and the epithelial surface is indicated by a white dashed line. Scale bar, 50 µm; n = 5 in each group. (F) Quantification of butyrate levels in the colonic luminal content of GF mice colonized with human microbiota from aged or young donors, as described in D. Statistical significance was determined by unpaired t -test; **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05. n = 5 per group. All error bars are SEM.

Article Snippet: Sections from formalin-fixed paraffin-embedded (FFPE) samples were used for immunofluorescence with primary antibodies against MUC2 (Santa Cruz Biotechnology #sc-7314, 1:50) and CA1 (Santa Cruz Biotechnology #sc-393490, 1:200).

Techniques: Staining, Immunofluorescence, Transplantation Assay

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