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: BMC Biology

Article Title: Ileum tissue single-cell mRNA sequencing elucidates the cellular architecture of pathophysiological changes associated with weaning in piglets

doi: 10.1186/s12915-022-01321-3

Figure Lengend Snippet: Heterogeneity of epithelial secretory cell subtypes in ileum tissues of piglets. a t-SNE plot displaying 832 epithelial secretory cells (ESCs) separated into 6 subtypes. EC, enterocyte; GC, goblet cell; EEC, enteroendocrine cell; B/OC, BEST4 / OTOP2 cell. b Violin plots showing canonical marker genes across ESC subtypes. See Additional file : Table S2 for all marker genes. c Heatmap reflecting differentially expressed transcription factors and cell-type-specific genes in ESC subtypes. d Immunofluorescence (IF) staining assay of GC marker MUC2 (green) and EEC markers CCK (red) and GCG (red) ( n =6); arrow, target cell type; scale bars, 50 μm. e Differentiation pseudotime trajectory analysis of ESC subtypes. Predicted secretory lineage cells include GC, EEC, and B/OC; enterocytes (EC) represent absorptive cells. Red arrow indicates the direction of differentiation. f The differentially expressed genes (rows) along the pseudotime (columns) of secretory and absorptive cells clustering hierarchically into four profiles. The representative gene functions and pathways of each profile are shown

Article Snippet: The primary antibodies were involved in IF staining: rabbit anti- MUC2 (Biorbyt Ltd., orb372331, 1:200); rabbit polyclonal cholecystokinin ( CCK ) antibody (Origene, #20078, 1:150); mouse anti- GCG monoclonal antibody (Novus Biologicals, NBP2-21803, 1:2000); rabbit anti -CD45 (intracellular domain) (Cell Signaling, #13917, 1:300); mouse anti-Sybdecan-1 (Abcam, #ab181789, 1:250); rabbit anti-IgA alpha chain (Abcam, #ab97000, 1:400); rabbit anti- CD3 (Abcam, #ab135372, 1:200); rabbit anti- TNF alpha (Abcam, #EPR19147, 1:1,000); rabbit anti -IL1β mAb (Cell Signaling, #D3U3E, 1:500); rabbit anti- BID (Abcam, #ab32060, 1:100); rabbit anti- CD3 -(E4T1B) XP® mAb (CST, #78588, 1:400); mouse anti-CD8α (RPA-T8) mAb (CST, #55397, 1:400); rabbit anti-granzyme B (D2H2F) mAb (CST, #17215, 1:800); rabbit anti-perforin (E9F7N) mAb (Cell Signaling, #44865, 1:100); and rabbit anti- ITGAE/CD103 (EP206) mAb (Cell Signaling, #95835, 1:150).

Techniques: Marker, Immunofluorescence, Staining

Journal: Scientific reports

Article Title: Robust and reproducible human intestinal organoid-derived monolayer model for analyzing drug absorption.

doi: 10.1038/s41598-025-95823-z

Figure Lengend Snippet: Fig. 6. Characterization of the novel human duodenal organoid-derived monolayer model. (a) Immunofluorescence analysis for E-cadherin (red), ZO-1 (green), (b) MUC2 (red), and nucleus (blue). Scale bars, 100 µm. (c) Immunofluorescence analysis depicting cross sections of the monolayer model; ZO-1 (green), nucleus (blue), and F-actin (white). Scale bars, 100 µm. Black arrows indicate the Transwell membrane. (d) and (e) Transmission electron microscopic analysis of the monolayer model. Mv, microvilli; Tj, tight junction; Sg, secretory granule-like structures. Scale bars, 10 µm for low magnification images and 2 µm for high magnification images. (f) Efflux transporter P-gp assay using the P-gp substrate digoxin with or without zosuquidar as an inhibitor. AtoB, apical to basal; BtoA, basal to apical. (g) Efflux transporter assay using BCRP substrate sulfasalazine with or without Ko143. (h) UDP-glucuronosyltransferase (UGT) functional assay using raloxifene as a UGT substrate. (i) Carboxylesterase 2 (CES2) functional assay using irinotecan as a substrate with or without loperamide as an inhibitor. The amount of SN-38, a metabolite of irinotecan, was detected in the cells of the monolayer model. (j) and (k) Midazolam permeability and metabolism assay using 2 µM midazolam as a substrate of CYP3A with or without 1-aminobenzotriazole (ABT) as a pan-CYP inhibitor. BLQ, below limit of quantification. (l) Relative gene expression determined by quantitative RT-PCR of the monolayer model with induction of VD3 or VD3 + rifampicin. Cells were treated with inducers for the last two days of the differentiation process. The P value was determined by a two-tailed unpaired t-test.

Article Snippet: The monolayer was incubated with primary antibodies [anti-ZO-1 antibody Alexa Fluor 488 (MA3-39,100-A488; Thermo Fisher Scientific), anti-E-cadherin antibody (PA5-32,178; Thermo Fisher Scientific), and anti-MUC2 antibody (NBP225,221; Novus Biologicals)] in blocking solution containing 0.3% Triton X-100 at 4 °C overnight.

Techniques: Derivative Assay, Immunofluorescence, Membrane, Transmission Assay, Functional Assay, Permeability, Gene Expression, Quantitative RT-PCR, Two Tailed Test

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Enhanced Utilization of Induced Pluripotent Stem Cell–Derived Human Intestinal Organoids Using Microengineered Chips

doi: 10.1016/j.jcmgh.2017.12.008

Figure Lengend Snippet: Table of Antibodies Used in Study

Article Snippet: Mucin-2 (CCP58) , 1:500 , NBP2-25221 , Mouse IgG1 , Novus Biologicals.

Techniques:

Journal: Immunology

Article Title: Human Gut Commensal Bacteroides fragilis Suppresses Mucin Production and Alters Microbiota Composition Resulting in Accelerated Type 1 Diabetes in Mice

doi: 10.1111/imm.70032

Figure Lengend Snippet: Suppressed mucin production in the distal colon of conventionalized BF‐colonised mice. Sections of formalin fixed distal colon and ileum tissues (with luminal contents) of 12‐week‐old conventionalized GF (GF‐conv/Ex‐GF) and conventionalized BF‐monocolonised (BF‐conv/Ex‐BF) mice were subjected to immunofluorescence staining using anti‐Muc2 antibody, or Alcian blue/nuclear fast, periodic acid‐Schiff's/haematoxylin, and Alcian blue periodic acid‐Schiff's/haematoxylin staining. (A) Representative images of immunofluorescence staining of colon and ileum tissue sections (left panels) and relative fluorescence values of Muc2 staining of multiple sections of distal colon tissues from 3 mice/group (right panel) are shown. (B) Representative images Alcian blue/nuclear fast, periodic acid‐Schiff's/haematoxylin, and Alcian blue periodic acid‐Schiff's/haematoxylin staining of colon and ileum tissue sections are shown.

Article Snippet: Tissue sections were stained using a mouse Muc2 specific antibody (Novus Biologicals) followed by an Alexaflor‐488 linked secondary antibody.

Techniques: Immunofluorescence, Staining, Fluorescence