Journal: Journal of Functional Foods

Article Title: Lactobacillus plantarum ZJUIDS04 alleviates DSS-induced colitis via modulating gut microbiota

doi: 10.1016/j.jff.2023.105794

Figure Lengend Snippet: Fig. 4. Effect of L. plantarum ZJUIDS04 on gut-barrier barrier function in DSS-induced colitis mice. (A) Expression of ZO-1, claudin-1, occludin, and in colon tissue evaluated by immunofluorescence staining. Scale bars, 100 μm. (B) Quantitative analysis of ZO-1, claudin-1, and occludin. (C) MUC2. (D) Relative expression of MUC2. All data are expressed as means ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001, vs the DSS group.

Article Snippet: Western blotting was performed as described (Ding et al., 2022) using the following antibodies: MUC2 (Affnity Biosciences, USA), and anti-β-actin (Boster C. Yu et al. Journal of Functional Foods 109 (2023) 105794 Biological Technology, Wuhan, China).

Techniques: Expressing, Immunofluorescence, Staining

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: Nutrients

Article Title: Metabolism of Caprine Milk Carbohydrates by Probiotic Bacteria and Caco-2:HT29–MTX Epithelial Co-Cultures and Their Impact on Intestinal Barrier Integrity

doi: 10.3390/nu10070949

Figure Lengend Snippet: Fold change of MUC4 , MUC2 , and MUC5AC mRNA from Caco-2:HT29–MTX (90:10) co-cultures after 3 h incubation with ( A ) probiotic bacteria or a carbohydrate fraction (CF) relative to untreated co-cultures; ( B ) bacteria/CF combinations compared to untreated monolayers; ( C ) bacteria/CF combinations compared to monolayers incubated with CF; and ( D ) bacteria/CF combinations relative to bacteria only controls. Data are expressed as the mean fold change (± SEM) of three replicates across three independent experiments; n = 3 A statistically significant difference in fold change at ±1.5 is indicated by * ( P < 0.05). LGG = L. rhamnosus GG; HN001 = L. rhamnosus HN001; Lp299v = L. plantarum 299v and LcS = L. casei Shirota.

Article Snippet: The abundance of mucin proteins in cell lysate (CL) and spent media (SM) was determined by indirect enzyme linked immunosorbent assay (indirect ELISA) using MUC2 mouse mono-clonal antibody (clone 4A4, 1:250; Creative Biomart, New York, NY, USA), MUC4 mouse mono-clonal antibody (clone 5B12, 1:500; Abnova, Taipei, Taiwan) or MUC5AC mouse mono-clonal antibody (clone 2H7, 1:250; Abnova) and horseradish peroxidase-rabbit anti-mouse immunoglobulin G conjugate (Abcam, Cambridge, UK; 1:5000 dilution for MUC2 and MUC5AC and 1:10,000 dilution for MUC4) as described previously [ ].

Techniques: Incubation, Bacteria

Journal: Nutrients

Article Title: Metabolism of Caprine Milk Carbohydrates by Probiotic Bacteria and Caco-2:HT29–MTX Epithelial Co-Cultures and Their Impact on Intestinal Barrier Integrity

doi: 10.3390/nu10070949

Figure Lengend Snippet: The abundance of MUC4, MUC2 and MUC5AC mucin protein from Caco-2:HT29–MTX (90:10) co-cultures after 3 h incubation with bacteria or a carbohydrate fraction (CF) from caprine milk either alone or in combination. Results are expressed as the mean abundance (±SEM); n = 3. * =significantly different ( P < 0.05) compared to untreated co-cultures, and z = significantly different ( P < 0.05) compared to co-cultures incubated with the respective bacteria only. UNT = untreated; LGG = L. rhamnosus GG; HN001 = L. rhamnosus HN001; Lp299v = L. plantarum 299v; and LcS = L. casei Shirota.

Article Snippet: The abundance of mucin proteins in cell lysate (CL) and spent media (SM) was determined by indirect enzyme linked immunosorbent assay (indirect ELISA) using MUC2 mouse mono-clonal antibody (clone 4A4, 1:250; Creative Biomart, New York, NY, USA), MUC4 mouse mono-clonal antibody (clone 5B12, 1:500; Abnova, Taipei, Taiwan) or MUC5AC mouse mono-clonal antibody (clone 2H7, 1:250; Abnova) and horseradish peroxidase-rabbit anti-mouse immunoglobulin G conjugate (Abcam, Cambridge, UK; 1:5000 dilution for MUC2 and MUC5AC and 1:10,000 dilution for MUC4) as described previously [ ].

Techniques: Incubation, Bacteria

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Antibodies Used in This Study

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Mouse TaqMan Gene Expression Assays Used in This Study

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Gene Expression

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Ectoine enhances production of secreted mucins in DS mice. ( A , C ) Representative images of IF staining showing the decreased production of MUC5AC ( A ) and MUC2 ( C ) in DS mice (DS+PBS) was recovered by ectoine eye drops (DS+Ect); Scale bars = 50 µm. The stitched images were made because the cornea and conjunctiva in some eyeball sections were too far apart to be captured on the same image. ( B , D ) IF intensity quantified the alternation of MUC5AC and MUC2 in 3 groups; each circle represents one eye. Data are shown as Mean ± SD, n = 6; the P values are shown in the graph as compared with the DS+PBS group.

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Staining, Eye Drops

Journal: Investigative Ophthalmology & Visual Science

Article Title: Ectoine Enhances Mucin Production Via Restoring IL-13/IFN-γ Balance in a Murine Dry Eye Model

doi: 10.1167/iovs.65.6.39

Figure Lengend Snippet: Gene expression of MUC5AC, MUC2, MUC1, MUC4, MUC16 , and MUC15 by conjunctiva and cornea. ( A ) Gene expression of all 6 mucins in normal mice were detected at different mRNA levels based on the threshold cycle (Ct) value in real-time PCR with house-keeping gene GAPDH as an internal control. ( B ) The mRNA levels of all six mucins in conjunctiva of the three groups of mice. ( C ) The mRNA expression of MUC1, MUC4, and MUC16 by corneas in the three groups of mice. Data are shown as Mean ± SD, n = 6; the P values are shown in the graph as compared with the DS+PBS group.

Article Snippet: MUC2 , CUSABIO , PA207869 , Rabbit , 1:3000.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control, Expressing

Journal: International Journal of Nanomedicine

Article Title: Oral Microcapsules Encapsulating Endometrial Regenerative Cell-Derived Exosomes Promote Intestinal Epithelial Barrier Repair and Ameliorate Experimental Colitis

doi: 10.2147/IJN.S540230

Figure Lengend Snippet: MFGE8 knockdown attenuates CSE-mediated restoration of the intestinal mucus barrier. ( A ) Representative Alcian blue-stained micrographs showing goblet cells (blue). Scale bar = 100 μm ( B ) Quantitative analysis of goblet cells in colon tissues. ( C ) MUC2 immunofluorescence staining (red, muc2 marker) and DAPI (blue, nuclear staining) in colonic tissues. Scale bar = 100 μm. ( D ) Statistical analysis of the mean fluorescence intensity of MUC2. ( E ) Immunofluorescence staining of CD11c (green, dendritic cell marker) and DAPI (blue, nuclear staining) in colonic tissues. Scale bar = 100 μm. ( F ) Integral optical density quantification of CD11c staining. Data were analyzed using one-way ANOVA; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Immunostaining was performed using a rabbit anti-MUC2 antibody (Affinity Biosciences).

Techniques: Knockdown, Staining, Immunofluorescence, Marker, Fluorescence