Journal: Gut

Article Title: CDX2 autoregulation in human intestinal metaplasia of the stomach: impact on the stability of the phenotype

doi: 10.1136/gut.2010.222323

Figure Lengend Snippet: Sequences of the oligonucleotides used for site-directed mutagenesis, RT-PCR, qPCR and chromatin immunoprecipitation (ChIP)

Article Snippet: MUC2 , TaqManGene Expression Assay MUC2, Hs03005094_m1.

Techniques: Mutagenesis, Chromatin Immunoprecipitation, Gene Expression, Expressing

Journal: Gut

Article Title: CDX2 autoregulation in human intestinal metaplasia of the stomach: impact on the stability of the phenotype

doi: 10.1136/gut.2010.222323

Figure Lengend Snippet: Effect of a dominant negative form of CDX2 on CDX2 targets and its endogenous expression. Fold increase of (A) MUC2, (B) LI-Cadherin and (C) endogeneous CDX2 mRNA in MKN45 and HCT116 cells transfected with CDX2 alone or together with a dominant negative form, evaluated by qPCR. The values obtained with mock-transfected cells were referred to as 1. mRNA levels were normalised to the corresponding TBP mRNA level.

Article Snippet: MUC2 , TaqManGene Expression Assay MUC2, Hs03005094_m1.

Techniques: Dominant Negative Mutation, Expressing, Transfection

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: Microbiome Research Reports

Article Title: A host-microbial metabolite interaction gut-on-a-chip model of the adult human intestine demonstrates beneficial effects upon inulin treatment of gut microbiome

doi: 10.20517/mrr.2023.79

Figure Lengend Snippet: Primer sequences

Article Snippet: MUC2 , TaqMan primer probes; Assay ID Hs03005103_g1 , .

Techniques:

Journal: Microbiome Research Reports

Article Title: A host-microbial metabolite interaction gut-on-a-chip model of the adult human intestine demonstrates beneficial effects upon inulin treatment of gut microbiome

doi: 10.20517/mrr.2023.79

Figure Lengend Snippet: mRNA expression levels of 11 genes determined by RT-qPCR in human colon tissue explants (donor 3) in the IEBC exposed to i-screen supernatant (untreated control, inulin-treated, and untreated control with added SCFA) for 24 h ( n = 5-6). (A) Tight junction complex genes OCLN, CLDN-1, CLDN-2, and ZO-1; (B) Mucus genes mucin MUC2 and mucin MUC5B; (C) Genes related to the inflammatory response: cytokine IL-8, TNFSF10, LBP, and CCL20; (D) Chromatin remodeler HDAC3. Target gene expression is expressed relative to the reference gene 36B4 and normalized to control. Error bars with points represent estimated marginal means with standard errors, obtained from linear mixed-effects models. * P < 0.05; ** P < 0.01. RT-qPCR: Reverse transcription quantitative PCR; IEBC: intestinal explant barrier chip; SCFA: short-chain fatty acid; OCLN: occludin; CLDN-1: claudin-1; CLDN-2: claudin-2; ZO-1: zonulin-1; IL-8: interleukin-8; TNFSF10: TNF superfamily member 10; LBP: LPS binding protein; CCL20: C-C motif chemokine ligand 20; HDAC3: histone deacetylase 3.

Article Snippet: MUC2 , TaqMan primer probes; Assay ID Hs03005103_g1 , .

Techniques: Expressing, Quantitative RT-PCR, Control, Targeted Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Binding Assay, Histone Deacetylase Assay