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: Scientific Reports

Article Title: Dietary polyamines promote intestinal adaptation in an experimental model of short bowel syndrome

doi: 10.1038/s41598-024-55258-4

Figure Lengend Snippet: Polyamines enhance mucosal defense factors in rats with massive intestinal resection. (A-B) Fecal and serum secretory IgA was measured using ELISA. n = 5–7/group. (C) IgA in ileum tissue was assessed using western blotting. n = 4–6/group. (D) Fecal mucin was measured using a fluorometric assay. n = 4–6/group. (E) Representative images of ileal villus showing immunostaining by Muc2 (original magnification, ×400; scale bars = 200 μm). Left graphs show the number of goblet cells per unit villous area and the size of goblet cell secretion granule. (F) The expression of Claudin-3 in the ileum tissue was measured by western blot analysis. Representative images of ileal villus showing immunostaining by Claudin-3 (original magnification, ×200; scale bars = 100 μm). (G) Serum DAO was measured by ELISA. n = 5–7/group. (H) Serum GLP-2 was measured by ELISA. n = 5–7/group. (I) Fecal short-chain fatty acid (SCFA) content was measured using high-performance liquid chromatography. n = 3–6/group. Data are presented as mean ± SD. Results of one-way ANOVA are represented as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Nonspecific antibody binding was blocked using 15% goat serum for 30 min. For staining of Ki-67, Muc2, and Claudin-3, the slides were then incubated with anti-Ki67 antibody (ab16667; Abcam, Cambridge, UK), anti-Muc2 rabbit monoclonal antibody (M01212-1; Boster Biological Technology, Pleasanton, CA, USA), and Claudin-3 polyclonal antibody (#PA5-32353; Invitrogen, Waltham, MA, USA) at 4°C overnight, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunostaining, Expressing, High Performance Liquid Chromatography

Journal: ACS chemical neuroscience

Article Title: Altered Secretion, Constitution, and Functional Properties of the Gastrointestinal Mucus in a Rat Model of Sporadic Alzheimer's Disease.

doi: 10.1021/acschemneuro.3c00223

Figure Lengend Snippet: Figure 3. Biochemical analysis of the GI mucus obtained from the rat model of sporadic AD induced by intracerebroventricular streptozotocin (STZ-icv) and the controls. (A) UV−vis spectra of mucus from the control and the STZ-icv animals (left) and the area under the curve reflecting dilution (right). (B) Absorbance of samples at 230, 260, and 280 nm, reflecting the concentration of salts, nucleic acids, and proteins. (C) Relationship between sample absorbance (log-transformed y-axis) and wavelength depicted for the demonstration of the relationship between the 400 nm peak and the UV region. (D) Model-derived estimates from the linear model reflecting the concentration of protein in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (E) Model-derived estimates from the linear model reflecting the signal intensity of AB; reflecting glycoprotein content) in the mucus of the STZ- icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (F) Mucin 2 (MUC2) dot blot, model-derived estimates from the linear model reflecting the concentration of MUC2 in the mucus of the STZ-icv and the controls (left) and the contrast illustrating the effect size (right). Mean estimates are accompanied by 95% confidence intervals. (G) Model- derived estimates from the linear model reflecting peak mastPASTA-obtained force (inversely correlated with lubrication capacity) in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (H) Model-derived estimates from the linear model reflecting the oxidation−reduction potential (ORP) in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (I) Model- derived estimates from the linear model reflecting the NRP-integrated density in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (J) Model-derived estimates from the linear model reflecting the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS)-derived reductive capacity (in equivalents of dithiothreitol [mM]) in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals. (J) Model-derived estimates from the linear model reflecting lipid peroxidation (estimated using the thiobarbituric acid reactive substances (TBARS) assay) in the mucus of the STZ-icv and the controls (upper) and the contrast illustrating the effect size (lower). Mean estimates are accompanied by 95% confidence intervals.

Article Snippet: Blocked membranes were incubated with the rabbit anti-MUC2 primary antibody (Boster Biological Technology, USA) diluted in the blocking buffer 500-fold for 24 h at 4 °C.

Techniques: Control, Concentration Assay, Transformation Assay, Derivative Assay, Dot Blot, TBARS Assay

Journal: Molecular Medicine

Article Title: Evaluation of two laboratory model methods for diarrheal irritable bowel syndrome

doi: 10.1186/s10020-022-00599-x

Figure Lengend Snippet: The primers used in this experiment

Article Snippet: The blocking solution is gently shaken off and a proportion of primary antibodies ZO-1 (BOSTER PB9234, Wuhan, China) and MUC2 (BOSTER BM5029, Wuhan, China) in PBS is added dropwise to the sections, which are incubated overnight at 4 °C in a wet box.

Techniques: Sequencing

Journal: Molecular Medicine

Article Title: Evaluation of two laboratory model methods for diarrheal irritable bowel syndrome

doi: 10.1186/s10020-022-00599-x

Figure Lengend Snippet: Intensity of ZO-1 and MUC2 (green) in ileal and colonic tissues of rats in each group. Nuclei stained with DAPI (blue) (200×) ( A ), mRNA expression levels of ZO-1 ( B ), MUC2 ( C ), OCLN ( D ), CLDN4 ( E ) in each group of rats.* p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The blocking solution is gently shaken off and a proportion of primary antibodies ZO-1 (BOSTER PB9234, Wuhan, China) and MUC2 (BOSTER BM5029, Wuhan, China) in PBS is added dropwise to the sections, which are incubated overnight at 4 °C in a wet box.

Techniques: Staining, Expressing

Journal: Pathogens and disease

Article Title: Bacillus subtilis RZ001 improves intestinal integrity and alleviates colitis by inhibiting the Notch signalling pathway and activating ATOH-1.

doi: 10.1093/femspd/ftaa016

Figure Lengend Snippet: Fig. 2 Effects of the bacterial strains on MUC2 expression. (A) Effect of the

Article Snippet: The supernatant was collected and then MUC2 levels were measured by a mouse MUC2 ELISA kit (CSB-E15065m, Cusabio, Wuhan, China).

Techniques: Expressing