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  • 92
    Thermo Fisher muc 2 antibody
    Western blot and dot blot analysis of colon differentiation and CSLCs markers following fatty acids treatments. ( A ) Total lysate were obtained from COLO 320 DM cells treated with vehicle control (CTRL VH), 25 uM EPA or SA for 48 and 96 hours. Western blot was performed to study the expression of cytokeratin-20 (CK20) and CD133 after treatment with the fatty acids. Dot blot was performed to study the expression of Mucin 2 (MUC2) after treatment with the fatty acids. A peptide identical to the one used to generate the <t>Muc-2</t> antibody was used in a peptide competition assay to determine MUC2 antibody specificity. The changes in ( B ) CD133, ( C ) CK20, and ( D ) MUC2 protein expression with respect to β-Actin are representative of three separate experiments. Results represent the mean ± SD of at least three experiments. p values were calculated with Student's t-test on treated samples vs. CTRL VH (* p≤0.05, ** p≤0.01).
    Muc 2 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    muc 2  (Abcam)
    95
    Abcam muc 2
    Muc-5ac expression in the stomach was not affected by indomethacin administration. Muc-5ac immunoreactivity was evident in the gastric mucosal of C57Bl6 and <t>Muc-2–deficient</t> ( Muc2 −/− ) mice, and was not changed by administration of indomethacin (10 mg/kg 12 hours before harvesting of tissue).
    Muc 2, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti muc 2 antibodies
    B. fragilis ZY-312 improves gut barrier integrity and function in the CDI mouse model. Representative images of hematoxylin-eosin stained cecum and colon tissue samples from all groups are shown. Images showing representative immunohistochemical staining of <t>Muc-2</t> and ZO-1 protein located in cecal and colon tissues in the Blank, CDI model, MTZ, and BFT groups are also shown. Mice in the Blank group were reared parallelly without any treatment. Mice in the CDI model group drank water with antibiotics including kanamycin (0.8 mg/mL), gentamicin (0.07 mg/mL), colistin (0.1135 mg/mL), metronidazole (0.43 mg/mL), and vancomycin (0.09 mg/mL) from D –9 to D –2, intraperitoneally injected with clindamycin (10 mg/kg) at D –1, and orally challenged with 3 × 10 8 cfu of C. difficile spores from D0 to D2. Mice in the MTZ group were treated with metronidazole (1 mg/day) from the day of C. difficile spore challenge (D0) to D5. Mice in the BFT groups were CDI model mice prophylactically treated with 1 × 10 7 or 1 × 10 8 cfu/day B. fragilis from the onset (D –9) to the end of CDI modeling (D2). Data of mean optical density are shown as means ± SEM. ∗ p
    Anti Muc 2 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology muc 2
    Ceftriaxone treatment results in alterations of intestinal innate barrier defenses. (A and B) qRT-PCR results of Paneth cell antimicrobials from DSI of saline- and ceftriaxone-treated mice colonized with EF CK135 and euthanized at day 4 after the first dose of ceftriaxone administration (A) and RegIIIγ at different time points (B). GAPDH was used as an internal control. CFU of EF CK135 in the intestinal sections (C) and in the liver, spleen, and MLN (D) of RegIIIγ −/− mice treated with saline or ceftriaxone and euthanized at day 4 after the first dose of ceftriaxone. (E) Enumeration of PAS/AB + goblet cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (F) Enumeration of <t>Muc2</t> + cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (G) Relative expression of Muc2 by qRT-PCR in the DSI and large intestinal sections from saline- and ceftriaxone-treated mice at day 4 after ceftriaxone administration. (H) Measurement of mucus layer thickness in the large intestines of saline- and ceftriaxone-treated mice at day 4 after ceftriaxone treatment. n = 5 mice per group. *, P values were
    Muc 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novocastra muc 2
    Immunohistchemical staining. CK7 ( A ) and Muc-5AC ( C ) were negative. CK20 ( B ) and <t>Muc-2</t> ( D ) were positive.
    Muc 2, supplied by Novocastra, used in various techniques. Bioz Stars score: 88/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Leica Microsystems muc 2
    Immunohistchemical staining. CK7 ( A ) and Muc-5AC ( C ) were negative. CK20 ( B ) and <t>Muc-2</t> ( D ) were positive.
    Muc 2, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology antihuman muc 2
    (A–F) Scaffolds with intestinal cells collected from different time points were sectioned in half along the longitudinal axis for immunostaining. Immunostaining of <t>MUC-2</t> and ZO-1of Caco-2/HT29-MTX cultured on scaffolds 1 week and 4 weeks post-bioreactor culture were imaged by confocal microscopy. MUC2 was visualized as red, ZO-1 as green, and DAPI as blue. (G–I) Intestinal epithelial cells on the luminal surface of scaffolds were detached, and total RNA was isolated for evaluation of gene expression levels of different biomarkers. Gene expression levels of intestinal epithelium biomarkers, ZO-1 (G), E-cadherin (H), and villin (I), were evaluated by quantitative reverse transcription–polymerase chain reaction (qRT-PCR) overtime in cultures. Data are presented as mean ± SEM, n = 3 in each group, * p
    Antihuman Muc 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher muc 2
    FEMI exposure results in decreased goblet and Paneth cell populations later in life. (A) Harvested intestinal tissue samples were quantified for goblet cell populations as a ratio of enterocytes. FEMI exposure caused significantly decreased goblet cell ratios compared to sham controls at P0 (FEMI 12% versus sham 16%, P =0.005, n =17), at P7 (FEMI 6.6% versus sham 9.9%, P =0.008, n =20) and at P28 (FEMI 8.2% versus sham 11%, P =0.04, n =25). (B) Gene expression of the goblet cell marker <t>Muc2</t> was quantified ( n =8 at each time point). FEMI significantly decreased Muc2 expression at P0 and P28 ( P =0.003 and 0.002 respectively). Muc2 expression was also significantly increased at P56, but the increase was less than a twofold change and was thus considered clinically non-significant (φ). (C) Tissues were also quantified for Paneth cell populations per intestinal crypt. FEMI exposure significantly decreased Paneth cell numbers at P28 (2.5 versus 4.1, P
    Muc 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology rabbit polyclonal muc 2
    FEMI exposure results in decreased goblet and Paneth cell populations later in life. (A) Harvested intestinal tissue samples were quantified for goblet cell populations as a ratio of enterocytes. FEMI exposure caused significantly decreased goblet cell ratios compared to sham controls at P0 (FEMI 12% versus sham 16%, P =0.005, n =17), at P7 (FEMI 6.6% versus sham 9.9%, P =0.008, n =20) and at P28 (FEMI 8.2% versus sham 11%, P =0.04, n =25). (B) Gene expression of the goblet cell marker <t>Muc2</t> was quantified ( n =8 at each time point). FEMI significantly decreased Muc2 expression at P0 and P28 ( P =0.003 and 0.002 respectively). Muc2 expression was also significantly increased at P56, but the increase was less than a twofold change and was thus considered clinically non-significant (φ). (C) Tissues were also quantified for Paneth cell populations per intestinal crypt. FEMI exposure significantly decreased Paneth cell numbers at P28 (2.5 versus 4.1, P
    Rabbit Polyclonal Muc 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Novus Biologicals muc2
    Milk-derived exosomes promote goblet cell expression in vivo . (A, B) Representative PAS stained histomicrographs and corresponding quantification of numbers of goblets cell per villus in the terminal ileum for the three experimental groups, control, NEC, and exosome-treated NEC mouse pups. (C, D) Representative micrographs for <t>MUC2</t> staining and corresponding quantification of MUC2+ goblet cells per villus for control, NEC, and exosome-treated NEC mouse pups. (E-F) Representative micrographs for GRP94 staining and quantification of GRP94+ cells per villus in each experimental group. Samples were taken from the terminal ileum of each group. Experiments were independently repeated 3 times with a total of nine mice per group. Data are presented as means ± SD. ***p
    Muc2, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Creative BioMart muc2
    Relative fold change of MUC mRNA from epithelial cell cultures incubated with 4 mg/mL caprine milk oligosaccharide-enriched fraction (CMOF). The expression of mucin genes MUC4 , <t>MUC2</t> and MUC5AC by 100:0, 90:10, 75:25, and 0:100 Caco-2:HT29-MTX cell cultures after 12 h incubation with 4.0 mg/mL CMOF, compared to respective control monolayers. The data are expressed as the mean fold change (±SEM) of three replicates across three independent experiments. A statistically significant difference in fold change is indicated by * ( p
    Muc2, supplied by Creative BioMart, used in various techniques. Bioz Stars score: 94/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti muc2
    The majority of Barrett’s oesophagus cells have a similar transcript profile to oesophageal submucosal gland (OSG) cells. a t-Distributed Stochastic Neighbour Embedding (t-SNE) plots of cells from all samples from four BO patients ( n = 1107 including brain control), showing similarity of cells in two dimensions, coloured by tissue type (yellow, oesophagus; green, gastric cardia; purple, duodenum; orange, Barrett’s oesophagus; pink, brain). Brain was used as a control. b t-SNE plot of cells from four BO patient samples ( A–D) , as in a ). c Sankey diagram showing how each tissue type sampled contributes to the clusters shown in b . Colours and labels on the left indicate sampled tissue (as in a ); colours and labels on the right indicate cluster (as in b ). d Mean BEARscc score for each grouping of ‘gland-like’ cells ( n = 372), which are a sub-set of gastric (G, n = 175), BO ( n = 78) and OSG cells ( n = 119): excluding gastric and BO cells that expressed CHGA or <t>MUC2</t> (to exclude enteroendocrine and goblet cells, respectively) and excluding oesophageal cells that did not express TFF3 (to exclude squamous cells). ‘Ensemble’ refers to all cells grouped together. Thresholds were set at the tenth centile of cells in which at least one transcript was detected from each gene of interest
    Anti Muc2, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 145 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology anti muc 2
    The majority of Barrett’s oesophagus cells have a similar transcript profile to oesophageal submucosal gland (OSG) cells. a t-Distributed Stochastic Neighbour Embedding (t-SNE) plots of cells from all samples from four BO patients ( n = 1107 including brain control), showing similarity of cells in two dimensions, coloured by tissue type (yellow, oesophagus; green, gastric cardia; purple, duodenum; orange, Barrett’s oesophagus; pink, brain). Brain was used as a control. b t-SNE plot of cells from four BO patient samples ( A–D) , as in a ). c Sankey diagram showing how each tissue type sampled contributes to the clusters shown in b . Colours and labels on the left indicate sampled tissue (as in a ); colours and labels on the right indicate cluster (as in b ). d Mean BEARscc score for each grouping of ‘gland-like’ cells ( n = 372), which are a sub-set of gastric (G, n = 175), BO ( n = 78) and OSG cells ( n = 119): excluding gastric and BO cells that expressed CHGA or <t>MUC2</t> (to exclude enteroendocrine and goblet cells, respectively) and excluding oesophageal cells that did not express TFF3 (to exclude squamous cells). ‘Ensemble’ refers to all cells grouped together. Thresholds were set at the tenth centile of cells in which at least one transcript was detected from each gene of interest
    Anti Muc 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Novus Biologicals muc2 antibody
    Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and <t>MUC2</t> ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).
    Muc2 Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology muc2 h
    Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and <t>MUC2</t> ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).
    Muc2 H, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Abcam mucin 2 muc2
    Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and <t>MUC2</t> ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).
    Mucin 2 Muc2, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher rabbit polyclonal muc2 antibody
    Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and <t>MUC2</t> ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).
    Rabbit Polyclonal Muc2 Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Western blot and dot blot analysis of colon differentiation and CSLCs markers following fatty acids treatments. ( A ) Total lysate were obtained from COLO 320 DM cells treated with vehicle control (CTRL VH), 25 uM EPA or SA for 48 and 96 hours. Western blot was performed to study the expression of cytokeratin-20 (CK20) and CD133 after treatment with the fatty acids. Dot blot was performed to study the expression of Mucin 2 (MUC2) after treatment with the fatty acids. A peptide identical to the one used to generate the Muc-2 antibody was used in a peptide competition assay to determine MUC2 antibody specificity. The changes in ( B ) CD133, ( C ) CK20, and ( D ) MUC2 protein expression with respect to β-Actin are representative of three separate experiments. Results represent the mean ± SD of at least three experiments. p values were calculated with Student's t-test on treated samples vs. CTRL VH (* p≤0.05, ** p≤0.01).

    Journal: PLoS ONE

    Article Title: Omega-3 Eicosapentaenoic Acid Decreases CD133 Colon Cancer Stem-Like Cell Marker Expression While Increasing Sensitivity to Chemotherapy

    doi: 10.1371/journal.pone.0069760

    Figure Lengend Snippet: Western blot and dot blot analysis of colon differentiation and CSLCs markers following fatty acids treatments. ( A ) Total lysate were obtained from COLO 320 DM cells treated with vehicle control (CTRL VH), 25 uM EPA or SA for 48 and 96 hours. Western blot was performed to study the expression of cytokeratin-20 (CK20) and CD133 after treatment with the fatty acids. Dot blot was performed to study the expression of Mucin 2 (MUC2) after treatment with the fatty acids. A peptide identical to the one used to generate the Muc-2 antibody was used in a peptide competition assay to determine MUC2 antibody specificity. The changes in ( B ) CD133, ( C ) CK20, and ( D ) MUC2 protein expression with respect to β-Actin are representative of three separate experiments. Results represent the mean ± SD of at least three experiments. p values were calculated with Student's t-test on treated samples vs. CTRL VH (* p≤0.05, ** p≤0.01).

    Article Snippet: A peptide competition assays was performed to verify the specificity of the Muc-2 antibody (Pierce, ThermoScientifics, Waltham, MA).

    Techniques: Western Blot, Dot Blot, Expressing, Competitive Binding Assay

    Muc-5ac expression in the stomach was not affected by indomethacin administration. Muc-5ac immunoreactivity was evident in the gastric mucosal of C57Bl6 and Muc-2–deficient ( Muc2 −/− ) mice, and was not changed by administration of indomethacin (10 mg/kg 12 hours before harvesting of tissue).

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Muc-5ac expression in the stomach was not affected by indomethacin administration. Muc-5ac immunoreactivity was evident in the gastric mucosal of C57Bl6 and Muc-2–deficient ( Muc2 −/− ) mice, and was not changed by administration of indomethacin (10 mg/kg 12 hours before harvesting of tissue).

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Expressing, Mouse Assay

    Expression of Muc-2 and Muc-5ac in mice with gastric ulcers induced by serosal application of acetic acid (10 days before harvesting of tissue). Muc-5ac was expressed in all four groups of mice, with no marked differences in intensity of staining. Muc-2 was expressed, albeit at a low level ( arrowheads ), in both male and female wild-type mice (C57Bl6), but not in the Muc-2–deficient mice. The Muc-2 staining was localized mainly to the epithelium at the ulcer margin.

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Expression of Muc-2 and Muc-5ac in mice with gastric ulcers induced by serosal application of acetic acid (10 days before harvesting of tissue). Muc-5ac was expressed in all four groups of mice, with no marked differences in intensity of staining. Muc-2 was expressed, albeit at a low level ( arrowheads ), in both male and female wild-type mice (C57Bl6), but not in the Muc-2–deficient mice. The Muc-2 staining was localized mainly to the epithelium at the ulcer margin.

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Expressing, Mouse Assay, Staining

    Healing and bacterial colonization of gastric ulcers induced by application of acetic acid to the serosal surface of the stomach. A: Healing of ulcers was significantly impaired in male Muc-2–deficient ( Muc2 −/− ) mice, compared with the wild-type controls (C57). The mean ulcer area was similar in the four groups at 3 days after acetic acid application, but by 1 week later (day 10), the ulcers had healed to a significantly greater extent in the wild-type mice (both sexes) and in the female Muc-2–deficient mice (* P

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Healing and bacterial colonization of gastric ulcers induced by application of acetic acid to the serosal surface of the stomach. A: Healing of ulcers was significantly impaired in male Muc-2–deficient ( Muc2 −/− ) mice, compared with the wild-type controls (C57). The mean ulcer area was similar in the four groups at 3 days after acetic acid application, but by 1 week later (day 10), the ulcers had healed to a significantly greater extent in the wild-type mice (both sexes) and in the female Muc-2–deficient mice (* P

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Mouse Assay

    Indomethacin induces expression of Muc-2 in the gastric mucosa. Muc-2 was expressed in the colon, but not the stomach, of vehicle-treated C57Bl6 mice. At 12 hours after oral administration of indomethacin (10 mg/kg), Muc-2 immunoreactivity was evident ( arrowheads ) in the gastric mucosa. No Muc-2 immunoreactivity was detected in the stomach of Muc-2–deficient mice ( Muc2 −/− ), nor in the isotype controls.

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Indomethacin induces expression of Muc-2 in the gastric mucosa. Muc-2 was expressed in the colon, but not the stomach, of vehicle-treated C57Bl6 mice. At 12 hours after oral administration of indomethacin (10 mg/kg), Muc-2 immunoreactivity was evident ( arrowheads ) in the gastric mucosa. No Muc-2 immunoreactivity was detected in the stomach of Muc-2–deficient mice ( Muc2 −/− ), nor in the isotype controls.

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Expressing, Mouse Assay

    Healing of indomethacin-induced gastric damage was delayed in Muc-2–deficient ( Muc2 −/− ) mice, compared with wild-type controls (C57Bl6). Although the extent of damage induced by oral administration of indomethacin (10 mg/kg) was similar in the two groups, the gastric damage score was significantly reduced in the wild-type mice at 24 and 48 hours after indomethacin administration (* P

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Healing of indomethacin-induced gastric damage was delayed in Muc-2–deficient ( Muc2 −/− ) mice, compared with wild-type controls (C57Bl6). Although the extent of damage induced by oral administration of indomethacin (10 mg/kg) was similar in the two groups, the gastric damage score was significantly reduced in the wild-type mice at 24 and 48 hours after indomethacin administration (* P

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Mouse Assay

    Reduced gastric expression of COX-2 in male Muc-2–deficient ( Muc2 −/− ) mice. A: Female Muc-2–deficient mice exhibited greater (* P

    Journal: The American Journal of Pathology

    Article Title: Muc-2-Deficient Mice Display a Sex-Specific, COX-2-Related Impairment of Gastric Mucosal Repair

    doi: 10.1016/j.ajpath.2010.11.048

    Figure Lengend Snippet: Reduced gastric expression of COX-2 in male Muc-2–deficient ( Muc2 −/− ) mice. A: Female Muc-2–deficient mice exhibited greater (* P

    Article Snippet: For histology, sections were processed by routine methods and stained with H & E. For immunofluorescence detection and localization of muc-2 and muc-5AC, the sections were subjected to heat-induced epitope retrieval (Target retrieval solution, pH 9.0; DakoCytomation, Glostrup, Denmark), blocked with 3% bovine serum albumin, and subsequently incubated with either a polyclonal antibody raised against Muc-2 (1:500 dilution; Ab76774; Abcam, Cambridge, MA), or a monoclonal antibody raised against Muc-5ca (1:500 dilution; clone 45M1; Abcam), overnight at 4°C.

    Techniques: Expressing, Mouse Assay

    Examples of MUC2 expression in mucinous (colloid) adenocarcinoma and CDX2-positive adenocarcinoma

    Journal: Human pathology

    Article Title: MUC2 Expression is an Adverse Prognostic Factor in Superficial Gastroesophageal Adenocarcinomas

    doi: 10.1016/j.humpath.2013.10.020

    Figure Lengend Snippet: Examples of MUC2 expression in mucinous (colloid) adenocarcinoma and CDX2-positive adenocarcinoma

    Article Snippet: Tumors that express MUC2 were classified as “intestinal” type because MUC2 is normally expressed in intestinal goblet cells.

    Techniques: Expressing

    MUC2 Expression (Intestinal Type) is Associated with Poor Prognosis in Submucosal (T1b) EAC

    Journal: Human pathology

    Article Title: MUC2 Expression is an Adverse Prognostic Factor in Superficial Gastroesophageal Adenocarcinomas

    doi: 10.1016/j.humpath.2013.10.020

    Figure Lengend Snippet: MUC2 Expression (Intestinal Type) is Associated with Poor Prognosis in Submucosal (T1b) EAC

    Article Snippet: Tumors that express MUC2 were classified as “intestinal” type because MUC2 is normally expressed in intestinal goblet cells.

    Techniques: Expressing

    Regulatory effect of melatonin on hypermethylation of Muc2 promoter induced by rVvpM. a Genomic DNA treated with rVvpM for 12 h was prepared. Time responses of rVvpM in methylation status of Muc2 gene at − 274 and − 193 CpG sites were determined by methyl-specific PCR (MSP) analysis. The relative level of Muc2 methylation is shown, compared to the unmethylated form. Data represent means ± S.E. n = 4. *, p

    Journal: Journal of Biomedical Science

    Article Title: Melatonin restores Muc2 depletion induced by V. vulnificus VvpM via melatonin receptor 2 coupling with Gαq

    doi: 10.1186/s12929-019-0606-x

    Figure Lengend Snippet: Regulatory effect of melatonin on hypermethylation of Muc2 promoter induced by rVvpM. a Genomic DNA treated with rVvpM for 12 h was prepared. Time responses of rVvpM in methylation status of Muc2 gene at − 274 and − 193 CpG sites were determined by methyl-specific PCR (MSP) analysis. The relative level of Muc2 methylation is shown, compared to the unmethylated form. Data represent means ± S.E. n = 4. *, p

    Article Snippet: The following antibodies were purchased: PKCδ antibody (BD Biosciences, Franklin Lakes, NJ, USA); p-ERK, ERK, p-PKC, PKC and β-actin antibodies (Santa Cruz Biotechnology, Paso Robles, CA, USA); Muc2 antibody (Abcam, Cambridge, MA, USA); and horseradish peroxidase (HRP)-conjugated goat anti-rabbit and goat anti-mouse IgG antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

    Techniques: Methylation, Polymerase Chain Reaction

    Melatonin regulates the level of Muc2 in intestinal epithelial cells treated with rVvpM. Dose ( a ) and time ( b ) responses of rVvpM in Mucin (Muc) 2 mRNAs of HT29-MTX cells are shown. Data represent means ± S.E. n = 5. *, p

    Journal: Journal of Biomedical Science

    Article Title: Melatonin restores Muc2 depletion induced by V. vulnificus VvpM via melatonin receptor 2 coupling with Gαq

    doi: 10.1186/s12929-019-0606-x

    Figure Lengend Snippet: Melatonin regulates the level of Muc2 in intestinal epithelial cells treated with rVvpM. Dose ( a ) and time ( b ) responses of rVvpM in Mucin (Muc) 2 mRNAs of HT29-MTX cells are shown. Data represent means ± S.E. n = 5. *, p

    Article Snippet: The following antibodies were purchased: PKCδ antibody (BD Biosciences, Franklin Lakes, NJ, USA); p-ERK, ERK, p-PKC, PKC and β-actin antibodies (Santa Cruz Biotechnology, Paso Robles, CA, USA); Muc2 antibody (Abcam, Cambridge, MA, USA); and horseradish peroxidase (HRP)-conjugated goat anti-rabbit and goat anti-mouse IgG antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

    Techniques:

    Melatonin restores Muc2 depletion induced by VvpM in mouse ileum infected with V.vulnificus . Mice inoculated with boiled V. vulnificus (Cont), V. vulnificus (WT), VvpM mutant (mut), and VvpM complement (comp) at 1.3 × 10 9 CFU/mL, and sacrificed 16 h later. Mice were also given oral administration of melatonin (10 mg/kg) for 7 days prior to infection of V. vulnificus (WT). a Expression of Muc2 in mouse ileum was examined by immunofluorescence (Top panel, green) and immunohistochemical analysis (bottom panel, brown). Propidium iodide (PI, red) was used for nuclear counterstaining for immunofluorescence analysis. Scale bars, 100 μm. The mean numbers of Muc2-labeled cells per villi are shown in the bar graph. Data represent means ± S.E. n = 8. *, p

    Journal: Journal of Biomedical Science

    Article Title: Melatonin restores Muc2 depletion induced by V. vulnificus VvpM via melatonin receptor 2 coupling with Gαq

    doi: 10.1186/s12929-019-0606-x

    Figure Lengend Snippet: Melatonin restores Muc2 depletion induced by VvpM in mouse ileum infected with V.vulnificus . Mice inoculated with boiled V. vulnificus (Cont), V. vulnificus (WT), VvpM mutant (mut), and VvpM complement (comp) at 1.3 × 10 9 CFU/mL, and sacrificed 16 h later. Mice were also given oral administration of melatonin (10 mg/kg) for 7 days prior to infection of V. vulnificus (WT). a Expression of Muc2 in mouse ileum was examined by immunofluorescence (Top panel, green) and immunohistochemical analysis (bottom panel, brown). Propidium iodide (PI, red) was used for nuclear counterstaining for immunofluorescence analysis. Scale bars, 100 μm. The mean numbers of Muc2-labeled cells per villi are shown in the bar graph. Data represent means ± S.E. n = 8. *, p

    Article Snippet: The following antibodies were purchased: PKCδ antibody (BD Biosciences, Franklin Lakes, NJ, USA); p-ERK, ERK, p-PKC, PKC and β-actin antibodies (Santa Cruz Biotechnology, Paso Robles, CA, USA); Muc2 antibody (Abcam, Cambridge, MA, USA); and horseradish peroxidase (HRP)-conjugated goat anti-rabbit and goat anti-mouse IgG antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

    Techniques: Infection, Mouse Assay, Mutagenesis, Expressing, Immunofluorescence, Immunohistochemistry, Labeling

    Effect of dietary fats on Muc2 expression in rat colon. A) Muc2 mRNA level that was quantified by RT‐qPCR, normalized to the housekeeping gene GAPDH and calculated as fold changes (2 −ΔΔCt ) in which the soybean oil group was set as control. B) Relative abundance of Muc2 protein that was quantified by western blotting and normalized to β‐actin. a, b, means with different letters differed significantly ( p

    Journal: Molecular Nutrition & Food Research

    Article Title: Intake of Fish Oil Specifically Modulates Colonic Muc2 Expression in Middle‐Aged Rats by Suppressing the Glycosylation Process

    doi: 10.1002/mnfr.201700661

    Figure Lengend Snippet: Effect of dietary fats on Muc2 expression in rat colon. A) Muc2 mRNA level that was quantified by RT‐qPCR, normalized to the housekeeping gene GAPDH and calculated as fold changes (2 −ΔΔCt ) in which the soybean oil group was set as control. B) Relative abundance of Muc2 protein that was quantified by western blotting and normalized to β‐actin. a, b, means with different letters differed significantly ( p

    Article Snippet: Electrophoresis was run at 120 V for 2 h at 4 °C (Bio‐Rad Mini‐Protean) and western blotting was performed with primary antibodies against Muc2 (1:4000; Abcam, Cambridge, UK) and β‐actin (1:3000; Bioworld, Nanjing, China).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    B. fragilis ZY-312 improves gut barrier integrity and function in the CDI mouse model. Representative images of hematoxylin-eosin stained cecum and colon tissue samples from all groups are shown. Images showing representative immunohistochemical staining of Muc-2 and ZO-1 protein located in cecal and colon tissues in the Blank, CDI model, MTZ, and BFT groups are also shown. Mice in the Blank group were reared parallelly without any treatment. Mice in the CDI model group drank water with antibiotics including kanamycin (0.8 mg/mL), gentamicin (0.07 mg/mL), colistin (0.1135 mg/mL), metronidazole (0.43 mg/mL), and vancomycin (0.09 mg/mL) from D –9 to D –2, intraperitoneally injected with clindamycin (10 mg/kg) at D –1, and orally challenged with 3 × 10 8 cfu of C. difficile spores from D0 to D2. Mice in the MTZ group were treated with metronidazole (1 mg/day) from the day of C. difficile spore challenge (D0) to D5. Mice in the BFT groups were CDI model mice prophylactically treated with 1 × 10 7 or 1 × 10 8 cfu/day B. fragilis from the onset (D –9) to the end of CDI modeling (D2). Data of mean optical density are shown as means ± SEM. ∗ p

    Journal: Frontiers in Microbiology

    Article Title: Bacteroides fragilis Prevents Clostridium difficile Infection in a Mouse Model by Restoring Gut Barrier and Microbiome Regulation

    doi: 10.3389/fmicb.2018.02976

    Figure Lengend Snippet: B. fragilis ZY-312 improves gut barrier integrity and function in the CDI mouse model. Representative images of hematoxylin-eosin stained cecum and colon tissue samples from all groups are shown. Images showing representative immunohistochemical staining of Muc-2 and ZO-1 protein located in cecal and colon tissues in the Blank, CDI model, MTZ, and BFT groups are also shown. Mice in the Blank group were reared parallelly without any treatment. Mice in the CDI model group drank water with antibiotics including kanamycin (0.8 mg/mL), gentamicin (0.07 mg/mL), colistin (0.1135 mg/mL), metronidazole (0.43 mg/mL), and vancomycin (0.09 mg/mL) from D –9 to D –2, intraperitoneally injected with clindamycin (10 mg/kg) at D –1, and orally challenged with 3 × 10 8 cfu of C. difficile spores from D0 to D2. Mice in the MTZ group were treated with metronidazole (1 mg/day) from the day of C. difficile spore challenge (D0) to D5. Mice in the BFT groups were CDI model mice prophylactically treated with 1 × 10 7 or 1 × 10 8 cfu/day B. fragilis from the onset (D –9) to the end of CDI modeling (D2). Data of mean optical density are shown as means ± SEM. ∗ p

    Article Snippet: Next, the cells were incubated with anti-ZO-1 or anti-Muc-2 antibodies (Abcam, Cambridge, United Kingdom) in PBST containing 1% BSA at 4°C overnight, washed three times with PBST, incubated with a secondary antibody conjugated with DyLight 488 (Abbkine, Wuhan, China) at room temperature for 1 h, washed three times with PBST, and incubated with 10 μg/ml DAPI for 10 min. After removal of the DAPI solution, one drop of fluorescence quenching agent was added to the cell surface and the cells were examined under a fluorescence microscope (TE2000-U; Nikon).

    Techniques: Staining, Immunohistochemistry, Mouse Assay, Injection

    Bacteroides fragilis ZY-312 increases expression of the tight junction protein ZO-1 and mucin MUC-2 in colon cells infected with C. difficile . Representative images for immunofluorescence staining of tight junction protein ZO-1 (top) in Caco-2 cells and mucin Muc-2 (middle) in HT-29 cells are shown for all groups. (A) Blank control group, 4 × 10 4 Caco-2 or HT-29 cells were cultured without treatment. (B) B. fragilis group, cells were incubated with 4 × 10 7 cfu B. fragilis . (C) C. difficile group, cells were incubated with 4 × 10 6 cfu C. difficile . (D) Exclusion group, cells were infected with 4 × 10 7 cfu B. fragilis for the first hour and 4 × 10 6 cfu C. difficile for the second hour. (E) Competition group, cells were co-infected with B. fragilis and C. difficile . (F) Substitution group, C. difficile was added for the first hour and B. fragilis added at the second hour. The cells were incubated at 37°C under anaerobic conditions for 2 h.

    Journal: Frontiers in Microbiology

    Article Title: Bacteroides fragilis Prevents Clostridium difficile Infection in a Mouse Model by Restoring Gut Barrier and Microbiome Regulation

    doi: 10.3389/fmicb.2018.02976

    Figure Lengend Snippet: Bacteroides fragilis ZY-312 increases expression of the tight junction protein ZO-1 and mucin MUC-2 in colon cells infected with C. difficile . Representative images for immunofluorescence staining of tight junction protein ZO-1 (top) in Caco-2 cells and mucin Muc-2 (middle) in HT-29 cells are shown for all groups. (A) Blank control group, 4 × 10 4 Caco-2 or HT-29 cells were cultured without treatment. (B) B. fragilis group, cells were incubated with 4 × 10 7 cfu B. fragilis . (C) C. difficile group, cells were incubated with 4 × 10 6 cfu C. difficile . (D) Exclusion group, cells were infected with 4 × 10 7 cfu B. fragilis for the first hour and 4 × 10 6 cfu C. difficile for the second hour. (E) Competition group, cells were co-infected with B. fragilis and C. difficile . (F) Substitution group, C. difficile was added for the first hour and B. fragilis added at the second hour. The cells were incubated at 37°C under anaerobic conditions for 2 h.

    Article Snippet: Next, the cells were incubated with anti-ZO-1 or anti-Muc-2 antibodies (Abcam, Cambridge, United Kingdom) in PBST containing 1% BSA at 4°C overnight, washed three times with PBST, incubated with a secondary antibody conjugated with DyLight 488 (Abbkine, Wuhan, China) at room temperature for 1 h, washed three times with PBST, and incubated with 10 μg/ml DAPI for 10 min. After removal of the DAPI solution, one drop of fluorescence quenching agent was added to the cell surface and the cells were examined under a fluorescence microscope (TE2000-U; Nikon).

    Techniques: Expressing, Infection, Immunofluorescence, Staining, Cell Culture, Incubation

    Ceftriaxone treatment results in alterations of intestinal innate barrier defenses. (A and B) qRT-PCR results of Paneth cell antimicrobials from DSI of saline- and ceftriaxone-treated mice colonized with EF CK135 and euthanized at day 4 after the first dose of ceftriaxone administration (A) and RegIIIγ at different time points (B). GAPDH was used as an internal control. CFU of EF CK135 in the intestinal sections (C) and in the liver, spleen, and MLN (D) of RegIIIγ −/− mice treated with saline or ceftriaxone and euthanized at day 4 after the first dose of ceftriaxone. (E) Enumeration of PAS/AB + goblet cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (F) Enumeration of Muc2 + cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (G) Relative expression of Muc2 by qRT-PCR in the DSI and large intestinal sections from saline- and ceftriaxone-treated mice at day 4 after ceftriaxone administration. (H) Measurement of mucus layer thickness in the large intestines of saline- and ceftriaxone-treated mice at day 4 after ceftriaxone treatment. n = 5 mice per group. *, P values were

    Journal: Infection and Immunity

    Article Title: Ceftriaxone Administration Disrupts Intestinal Homeostasis, Mediating Noninflammatory Proliferation and Dissemination of Commensal Enterococci

    doi: 10.1128/IAI.00674-18

    Figure Lengend Snippet: Ceftriaxone treatment results in alterations of intestinal innate barrier defenses. (A and B) qRT-PCR results of Paneth cell antimicrobials from DSI of saline- and ceftriaxone-treated mice colonized with EF CK135 and euthanized at day 4 after the first dose of ceftriaxone administration (A) and RegIIIγ at different time points (B). GAPDH was used as an internal control. CFU of EF CK135 in the intestinal sections (C) and in the liver, spleen, and MLN (D) of RegIIIγ −/− mice treated with saline or ceftriaxone and euthanized at day 4 after the first dose of ceftriaxone. (E) Enumeration of PAS/AB + goblet cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (F) Enumeration of Muc2 + cells per 10 villi in the distal small intestinal tissue sections collected from saline- and ceftriaxone-treated mice. (G) Relative expression of Muc2 by qRT-PCR in the DSI and large intestinal sections from saline- and ceftriaxone-treated mice at day 4 after ceftriaxone administration. (H) Measurement of mucus layer thickness in the large intestines of saline- and ceftriaxone-treated mice at day 4 after ceftriaxone treatment. n = 5 mice per group. *, P values were

    Article Snippet: Tissues were then stained with murine Muc2 (H-300; Santa Cruz Biotechnology) antibody at 4°C for 90 min, followed by incubation with a goat anti-rabbit secondary antibody, staining with DAB for 2 min, and counterstaining with hematoxylin.

    Techniques: Quantitative RT-PCR, Mouse Assay, Expressing

    Immunohistchemical staining. CK7 ( A ) and Muc-5AC ( C ) were negative. CK20 ( B ) and Muc-2 ( D ) were positive.

    Journal: Journal of Ovarian Research

    Article Title: Mucinous adenocarcinoma of the intestinal type arising from mature cystic teratoma of the ovary: a rare case report and review of the literature

    doi: 10.1186/1757-2215-5-41

    Figure Lengend Snippet: Immunohistchemical staining. CK7 ( A ) and Muc-5AC ( C ) were negative. CK20 ( B ) and Muc-2 ( D ) were positive.

    Article Snippet: The immunohistochemical analysis showed positive staining for CK20 and Muc-2, while CK7, Muc-5AC and Muc-6 were negative in the adenocarcinomatous part.

    Techniques: Staining

    (A–F) Scaffolds with intestinal cells collected from different time points were sectioned in half along the longitudinal axis for immunostaining. Immunostaining of MUC-2 and ZO-1of Caco-2/HT29-MTX cultured on scaffolds 1 week and 4 weeks post-bioreactor culture were imaged by confocal microscopy. MUC2 was visualized as red, ZO-1 as green, and DAPI as blue. (G–I) Intestinal epithelial cells on the luminal surface of scaffolds were detached, and total RNA was isolated for evaluation of gene expression levels of different biomarkers. Gene expression levels of intestinal epithelium biomarkers, ZO-1 (G), E-cadherin (H), and villin (I), were evaluated by quantitative reverse transcription–polymerase chain reaction (qRT-PCR) overtime in cultures. Data are presented as mean ± SEM, n = 3 in each group, * p

    Journal: ACS Biomaterials Science & Engineering

    Article Title: Multifunctional Bioreactor System for Human Intestine Tissues

    doi: 10.1021/acsbiomaterials.7b00794

    Figure Lengend Snippet: (A–F) Scaffolds with intestinal cells collected from different time points were sectioned in half along the longitudinal axis for immunostaining. Immunostaining of MUC-2 and ZO-1of Caco-2/HT29-MTX cultured on scaffolds 1 week and 4 weeks post-bioreactor culture were imaged by confocal microscopy. MUC2 was visualized as red, ZO-1 as green, and DAPI as blue. (G–I) Intestinal epithelial cells on the luminal surface of scaffolds were detached, and total RNA was isolated for evaluation of gene expression levels of different biomarkers. Gene expression levels of intestinal epithelium biomarkers, ZO-1 (G), E-cadherin (H), and villin (I), were evaluated by quantitative reverse transcription–polymerase chain reaction (qRT-PCR) overtime in cultures. Data are presented as mean ± SEM, n = 3 in each group, * p

    Article Snippet: All specimens were then permeabilized using 0.1% Triton x-100 in phosphate-buffered saline (PBS, Gibco), then blocked with 5% bovine serum albumin (BSA, Sigma) for 2 h. These specimens were incubated overnight at 4 °C with antihuman ZO-1 (BD Transduction Laboratorie, 1:50), anti-e-cadherin (abcam, 1:50), antihuman-MUC-2 (Santa Cruz Biotech, 1:50), and anti-villin (abcam, 1:100), then immersed in Alexa Fluor 488 donkey antimouse and Alexa Fluor 546 goat–antirabbit secondary antibodies (Invitrogen) at a dilution of 1:100.

    Techniques: Immunostaining, Cell Culture, Confocal Microscopy, Isolation, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    Analysis of mucus thickness generated in the static culture and bioreactor cultures with oxygen control. (A) The quantification of mucus thickness. (B–G) Confocal images of MUC-2 staining on samples from different culture conditions. Scale bar = 10 μm. Data are presented as mean ± SEM, n = 5 in each group, * p

    Journal: ACS Biomaterials Science & Engineering

    Article Title: Multifunctional Bioreactor System for Human Intestine Tissues

    doi: 10.1021/acsbiomaterials.7b00794

    Figure Lengend Snippet: Analysis of mucus thickness generated in the static culture and bioreactor cultures with oxygen control. (A) The quantification of mucus thickness. (B–G) Confocal images of MUC-2 staining on samples from different culture conditions. Scale bar = 10 μm. Data are presented as mean ± SEM, n = 5 in each group, * p

    Article Snippet: All specimens were then permeabilized using 0.1% Triton x-100 in phosphate-buffered saline (PBS, Gibco), then blocked with 5% bovine serum albumin (BSA, Sigma) for 2 h. These specimens were incubated overnight at 4 °C with antihuman ZO-1 (BD Transduction Laboratorie, 1:50), anti-e-cadherin (abcam, 1:50), antihuman-MUC-2 (Santa Cruz Biotech, 1:50), and anti-villin (abcam, 1:100), then immersed in Alexa Fluor 488 donkey antimouse and Alexa Fluor 546 goat–antirabbit secondary antibodies (Invitrogen) at a dilution of 1:100.

    Techniques: Generated, Staining

    The differentiation of 3D intestinal epithelia. (a, b) General seeding cell seeding strategy for hInEpiC-derived and cell line-derived 3D intestinal constructs. (c-n) Immunohistological stainings of SI (sucrose-isomaltase, c, g, k), MUC-2 (Mucin 2, d, h, l), Lysozyme (e, i, m) and ChgA (Chromogranin A, f, g, n) showed the location of enterocytes, Goblet cells, Paneth cells, and enteroendocrine cells in differentiated HIE-derived, hInEpiC-derived cell line-derived epithelia. Scale bar, 25μm. (o) Relative mRNA expression levels of different markers of differentiated intestinal epithelia derived from the three different cell sources. The fold-change in mRNA expression was compared with cell line-derived 3D constructs at day 1 post cell seeding.

    Journal: PLoS ONE

    Article Title: In vitro enteroid-derived three-dimensional tissue model of human small intestinal epithelium with innate immune responses

    doi: 10.1371/journal.pone.0187880

    Figure Lengend Snippet: The differentiation of 3D intestinal epithelia. (a, b) General seeding cell seeding strategy for hInEpiC-derived and cell line-derived 3D intestinal constructs. (c-n) Immunohistological stainings of SI (sucrose-isomaltase, c, g, k), MUC-2 (Mucin 2, d, h, l), Lysozyme (e, i, m) and ChgA (Chromogranin A, f, g, n) showed the location of enterocytes, Goblet cells, Paneth cells, and enteroendocrine cells in differentiated HIE-derived, hInEpiC-derived cell line-derived epithelia. Scale bar, 25μm. (o) Relative mRNA expression levels of different markers of differentiated intestinal epithelia derived from the three different cell sources. The fold-change in mRNA expression was compared with cell line-derived 3D constructs at day 1 post cell seeding.

    Article Snippet: These specimens were incubated overnight at 4°C with anti-human ZO-1(1:100, BD Transduction Laboratories), anti-human-SI (sucrase-isomaltase) (1:100, Santa Cruz Biotech), anti-human-Muc-2 (1:50, Santa Cruz), anti-human-Lyz (1:100, Lysozyme, Abcam) and anti-CghA (1:100, Chromogranin A, Abcam, 1:100), then immersed in Alexa Fluor 488 donkey anti-mouse and Alexa Fluor 546 goat-anti-rabbit secondary antibodies (Invitrogen) at a dilution of 1:250, respectively.

    Techniques: Derivative Assay, Construct, Expressing

    FEMI exposure results in decreased goblet and Paneth cell populations later in life. (A) Harvested intestinal tissue samples were quantified for goblet cell populations as a ratio of enterocytes. FEMI exposure caused significantly decreased goblet cell ratios compared to sham controls at P0 (FEMI 12% versus sham 16%, P =0.005, n =17), at P7 (FEMI 6.6% versus sham 9.9%, P =0.008, n =20) and at P28 (FEMI 8.2% versus sham 11%, P =0.04, n =25). (B) Gene expression of the goblet cell marker Muc2 was quantified ( n =8 at each time point). FEMI significantly decreased Muc2 expression at P0 and P28 ( P =0.003 and 0.002 respectively). Muc2 expression was also significantly increased at P56, but the increase was less than a twofold change and was thus considered clinically non-significant (φ). (C) Tissues were also quantified for Paneth cell populations per intestinal crypt. FEMI exposure significantly decreased Paneth cell numbers at P28 (2.5 versus 4.1, P

    Journal: Disease Models & Mechanisms

    Article Title: Fetal exposure to maternal inflammation interrupts murine intestinal development and increases susceptibility to neonatal intestinal injury

    doi: 10.1242/dmm.040808

    Figure Lengend Snippet: FEMI exposure results in decreased goblet and Paneth cell populations later in life. (A) Harvested intestinal tissue samples were quantified for goblet cell populations as a ratio of enterocytes. FEMI exposure caused significantly decreased goblet cell ratios compared to sham controls at P0 (FEMI 12% versus sham 16%, P =0.005, n =17), at P7 (FEMI 6.6% versus sham 9.9%, P =0.008, n =20) and at P28 (FEMI 8.2% versus sham 11%, P =0.04, n =25). (B) Gene expression of the goblet cell marker Muc2 was quantified ( n =8 at each time point). FEMI significantly decreased Muc2 expression at P0 and P28 ( P =0.003 and 0.002 respectively). Muc2 expression was also significantly increased at P56, but the increase was less than a twofold change and was thus considered clinically non-significant (φ). (C) Tissues were also quantified for Paneth cell populations per intestinal crypt. FEMI exposure significantly decreased Paneth cell numbers at P28 (2.5 versus 4.1, P

    Article Snippet: qRT-PCR Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was performed using Taqman Fast Universal PCR Master Mix (2×) (Life Technologies) and Taqman Gene Expression Assays for Muc2 , Defa1 , cleaved caspase 3 and Ki67 (Life Technologies). qRT-PCR reactions were run in a C1000 Thermal Cycler (Eppendorf) and using the CFX96 Real-Time PCR Detection System (Bio-Rad).

    Techniques: Expressing, Marker

    Milk-derived exosomes promote goblet cell expression in vivo . (A, B) Representative PAS stained histomicrographs and corresponding quantification of numbers of goblets cell per villus in the terminal ileum for the three experimental groups, control, NEC, and exosome-treated NEC mouse pups. (C, D) Representative micrographs for MUC2 staining and corresponding quantification of MUC2+ goblet cells per villus for control, NEC, and exosome-treated NEC mouse pups. (E-F) Representative micrographs for GRP94 staining and quantification of GRP94+ cells per villus in each experimental group. Samples were taken from the terminal ileum of each group. Experiments were independently repeated 3 times with a total of nine mice per group. Data are presented as means ± SD. ***p

    Journal: PLoS ONE

    Article Title: Bovine milk-derived exosomes enhance goblet cell activity and prevent the development of experimental necrotizing enterocolitis

    doi: 10.1371/journal.pone.0211431

    Figure Lengend Snippet: Milk-derived exosomes promote goblet cell expression in vivo . (A, B) Representative PAS stained histomicrographs and corresponding quantification of numbers of goblets cell per villus in the terminal ileum for the three experimental groups, control, NEC, and exosome-treated NEC mouse pups. (C, D) Representative micrographs for MUC2 staining and corresponding quantification of MUC2+ goblet cells per villus for control, NEC, and exosome-treated NEC mouse pups. (E-F) Representative micrographs for GRP94 staining and quantification of GRP94+ cells per villus in each experimental group. Samples were taken from the terminal ileum of each group. Experiments were independently repeated 3 times with a total of nine mice per group. Data are presented as means ± SD. ***p

    Article Snippet: Of note, administration of milk-derived exosomes to the breastfed control mice did not show significant improvement to the MUC2 or GRP94 expression levels.

    Techniques: Derivative Assay, Expressing, In Vivo, Staining, Mouse Assay

    Milk-derived exosomes promote goblet cell expression in vitro . Exosomal protein (A) and RNA (B) content were taken up by LS174T cells. (C) Blue Periodic Acid Schiff (PAS) staining measuring the mucin production in LS174T control and exosome-treated cells (black arrows show positive stain). (D) Representative micrographs for MUC2 immunofluorescent staining of treatment and control groups (white arrows show positive stain). (E, F) Relative gene expression of goblet cell expression markers TFF3 , and MUC2 in both groups. (G) Representative immunofluorescent micrographs for GRP94 and (H) GRP94 gene expression in control and exosome-treated cells. Experiments were independently repeated 3 times. Data are presented as means ± SD. *p

    Journal: PLoS ONE

    Article Title: Bovine milk-derived exosomes enhance goblet cell activity and prevent the development of experimental necrotizing enterocolitis

    doi: 10.1371/journal.pone.0211431

    Figure Lengend Snippet: Milk-derived exosomes promote goblet cell expression in vitro . Exosomal protein (A) and RNA (B) content were taken up by LS174T cells. (C) Blue Periodic Acid Schiff (PAS) staining measuring the mucin production in LS174T control and exosome-treated cells (black arrows show positive stain). (D) Representative micrographs for MUC2 immunofluorescent staining of treatment and control groups (white arrows show positive stain). (E, F) Relative gene expression of goblet cell expression markers TFF3 , and MUC2 in both groups. (G) Representative immunofluorescent micrographs for GRP94 and (H) GRP94 gene expression in control and exosome-treated cells. Experiments were independently repeated 3 times. Data are presented as means ± SD. *p

    Article Snippet: Of note, administration of milk-derived exosomes to the breastfed control mice did not show significant improvement to the MUC2 or GRP94 expression levels.

    Techniques: Derivative Assay, Expressing, In Vitro, Staining

    Relative fold change of MUC mRNA from epithelial cell cultures incubated with 4 mg/mL caprine milk oligosaccharide-enriched fraction (CMOF). The expression of mucin genes MUC4 , MUC2 and MUC5AC by 100:0, 90:10, 75:25, and 0:100 Caco-2:HT29-MTX cell cultures after 12 h incubation with 4.0 mg/mL CMOF, compared to respective control monolayers. The data are expressed as the mean fold change (±SEM) of three replicates across three independent experiments. A statistically significant difference in fold change is indicated by * ( p

    Journal: Nutrients

    Article Title: Effect of a Semi-Purified Oligosaccharide-Enriched Fraction from Caprine Milk on Barrier Integrity and Mucin Production of Co-Culture Models of the Small and Large Intestinal Epithelium

    doi: 10.3390/nu8050267

    Figure Lengend Snippet: Relative fold change of MUC mRNA from epithelial cell cultures incubated with 4 mg/mL caprine milk oligosaccharide-enriched fraction (CMOF). The expression of mucin genes MUC4 , MUC2 and MUC5AC by 100:0, 90:10, 75:25, and 0:100 Caco-2:HT29-MTX cell cultures after 12 h incubation with 4.0 mg/mL CMOF, compared to respective control monolayers. The data are expressed as the mean fold change (±SEM) of three replicates across three independent experiments. A statistically significant difference in fold change is indicated by * ( p

    Article Snippet: In the colon adenocarcinoma LS174T cell line, galacto-oligosaccharides (8.0 mg/mL) stimulates the increased expression of MUC2 but not as a consequence of intracellular signalling through an NFĸB-mediated inflammatory cascade but rather as a consequence of direct interaction with goblet cells through an unknown mechanism [ ].

    Techniques: Incubation, Expressing

    The abundance of the individual mucin proteins (MUC4, MUC2 and MUC5AC) and the total mucin protein abundance from post-confluent (21 days post-seeding) control Caco-2:HT29-MTX mono- and co-cultures (ng/µg total protein) and those incubated with 4.0 mg/mL caprine milk oligosaccharide-enriched fraction (CMOF) for 12 h. Results are expressed as the mean abundance (ng/µg of total protein) (±SEM). * p

    Journal: Nutrients

    Article Title: Effect of a Semi-Purified Oligosaccharide-Enriched Fraction from Caprine Milk on Barrier Integrity and Mucin Production of Co-Culture Models of the Small and Large Intestinal Epithelium

    doi: 10.3390/nu8050267

    Figure Lengend Snippet: The abundance of the individual mucin proteins (MUC4, MUC2 and MUC5AC) and the total mucin protein abundance from post-confluent (21 days post-seeding) control Caco-2:HT29-MTX mono- and co-cultures (ng/µg total protein) and those incubated with 4.0 mg/mL caprine milk oligosaccharide-enriched fraction (CMOF) for 12 h. Results are expressed as the mean abundance (ng/µg of total protein) (±SEM). * p

    Article Snippet: In the colon adenocarcinoma LS174T cell line, galacto-oligosaccharides (8.0 mg/mL) stimulates the increased expression of MUC2 but not as a consequence of intracellular signalling through an NFĸB-mediated inflammatory cascade but rather as a consequence of direct interaction with goblet cells through an unknown mechanism [ ].

    Techniques: Incubation

    The majority of Barrett’s oesophagus cells have a similar transcript profile to oesophageal submucosal gland (OSG) cells. a t-Distributed Stochastic Neighbour Embedding (t-SNE) plots of cells from all samples from four BO patients ( n = 1107 including brain control), showing similarity of cells in two dimensions, coloured by tissue type (yellow, oesophagus; green, gastric cardia; purple, duodenum; orange, Barrett’s oesophagus; pink, brain). Brain was used as a control. b t-SNE plot of cells from four BO patient samples ( A–D) , as in a ). c Sankey diagram showing how each tissue type sampled contributes to the clusters shown in b . Colours and labels on the left indicate sampled tissue (as in a ); colours and labels on the right indicate cluster (as in b ). d Mean BEARscc score for each grouping of ‘gland-like’ cells ( n = 372), which are a sub-set of gastric (G, n = 175), BO ( n = 78) and OSG cells ( n = 119): excluding gastric and BO cells that expressed CHGA or MUC2 (to exclude enteroendocrine and goblet cells, respectively) and excluding oesophageal cells that did not express TFF3 (to exclude squamous cells). ‘Ensemble’ refers to all cells grouped together. Thresholds were set at the tenth centile of cells in which at least one transcript was detected from each gene of interest

    Journal: Nature Communications

    Article Title: Single cell RNA-seq reveals profound transcriptional similarity between Barrett’s oesophagus and oesophageal submucosal glands

    doi: 10.1038/s41467-018-06796-9

    Figure Lengend Snippet: The majority of Barrett’s oesophagus cells have a similar transcript profile to oesophageal submucosal gland (OSG) cells. a t-Distributed Stochastic Neighbour Embedding (t-SNE) plots of cells from all samples from four BO patients ( n = 1107 including brain control), showing similarity of cells in two dimensions, coloured by tissue type (yellow, oesophagus; green, gastric cardia; purple, duodenum; orange, Barrett’s oesophagus; pink, brain). Brain was used as a control. b t-SNE plot of cells from four BO patient samples ( A–D) , as in a ). c Sankey diagram showing how each tissue type sampled contributes to the clusters shown in b . Colours and labels on the left indicate sampled tissue (as in a ); colours and labels on the right indicate cluster (as in b ). d Mean BEARscc score for each grouping of ‘gland-like’ cells ( n = 372), which are a sub-set of gastric (G, n = 175), BO ( n = 78) and OSG cells ( n = 119): excluding gastric and BO cells that expressed CHGA or MUC2 (to exclude enteroendocrine and goblet cells, respectively) and excluding oesophageal cells that did not express TFF3 (to exclude squamous cells). ‘Ensemble’ refers to all cells grouped together. Thresholds were set at the tenth centile of cells in which at least one transcript was detected from each gene of interest

    Article Snippet: Sections were then blocked with normal goat serum (at room temperature) and incubated overnight at 4 °C with a primary antibody against anti-KRT14 (IHC, 1:1000, rabbit polyclonal, cat. PRB-155P, BioLegend), anti-TFF3 (IHC, 1:1000, mouse monoclonal, cat. WH0007033M1, Sigma-Aldrich®) , anti-MUC2 (IHC, 1:300, rabbit polyclonal, cat. SC-15334, Santa Cruz Biotechnology) , anti-CHGA (IHC, 1:500, rabbit polyclonal, cat. ab15160, Abcam) , anti-KRT7 (IHC, 1:4000, rabbit monoclonal, cat. ab181598, Abcam) , anti-LEFTY1 (IHC, 1:1000, D7E3G rabbit polyclonal, cat. 12647, Cell Signalling), anti-OLFM4 (IHC, 1:200, D1E4M rabbit monoclonal, cat. 14369, Cell Signalling Technology®), anti-ITLN1 (IHC/IF, 1:500, sheep polyclonal, cat. AF4254, R & D systems) , anti-MUC2 (IF, 1:300, mouse monoclonal, cat. ab11197, Abcam) or anti-SPINK4 (IF, 1:500, rabbit polyclonal, cat. HPA007286, Sigma-Aldrich®) .

    Techniques:

    Effect of S. flexneri infection on mucus expression. Enteroid monolayers derived from the colon were infected either apically or basolaterally with 4 × 10 7 CFU of WT S. flexneri for 2 h. Following invasion, the enteroid monolayers were incubated in medium containing gentamicin for an additional 4 h to allow intracellular replication of bacteria and to remove extracellular bacteria. Undifferentiated (ND) and differentiated monolayers (both uninfected) were used as controls. Whole-cell lysates of uninfected or infected enteroids were probed for Muc2 by Western blotting. A Western blot with β-actin is shown to confirm equal loading of samples (bottom panel).

    Journal: Infection and Immunity

    Article Title: Evaluating Shigella flexneri Pathogenesis in the Human Enteroid Model

    doi: 10.1128/IAI.00740-18

    Figure Lengend Snippet: Effect of S. flexneri infection on mucus expression. Enteroid monolayers derived from the colon were infected either apically or basolaterally with 4 × 10 7 CFU of WT S. flexneri for 2 h. Following invasion, the enteroid monolayers were incubated in medium containing gentamicin for an additional 4 h to allow intracellular replication of bacteria and to remove extracellular bacteria. Undifferentiated (ND) and differentiated monolayers (both uninfected) were used as controls. Whole-cell lysates of uninfected or infected enteroids were probed for Muc2 by Western blotting. A Western blot with β-actin is shown to confirm equal loading of samples (bottom panel).

    Article Snippet: Antibodies used were anti-Muc2 antibody (1:1,000) (Abcam) or anti-β-actin (1:5,000) (Invitrogen) diluted in 10% nonfat milk in PBS.

    Techniques: Infection, Expressing, Derivative Assay, Incubation, Western Blot

    Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and MUC2 ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).

    Journal: Acta Histochemica et Cytochemica

    Article Title: Reactivation of CDX2 in Gastric Cancer as Mark for Gene Silencing Memory

    doi: 10.1267/ahc.15014

    Figure Lengend Snippet: Wel glands in the deep invasive area ( A , HE, ×100) show negative for CDX2 ( B , ×100) and MUC2 ( C , ×100, and E , ×400, hatched area of C ) immunoreactivity, where MeCP2 is strongly positive in the nuclei of the invasive wel glands ( D , ×100, and F , ×400, hatched area of D ).

    Article Snippet: The specimens were then incubated with 2% non-fat dry milk in phosphate-buffered saline (PBS) for 10 min and with primary antibodies against CDX2 (M3636, DAKO, Tokyo, Japan), MUC2 (NBP1-31231, Novus Biologicals, USA, CO) and MeCP2 (D4F3, Cell Signaling Technology, Inc., USA, MA) for 15 min. After three 10-min washes with PBS, the specimens were incubated with rabbit anti-mouse IgG antibody preabsorbed with non-immunized serum.

    Techniques:

    Proper gastric mucosae of fundic gland area ( A , HE, ×100) show negative MUC2 ( B , ×100) and CDX2 ( C , ×100 and E , ×400, hatched areas of C ) expression. MeCP2 expression is, albeit less extensive than surrounding inflammatory cells, positive in the nuclei of proper gastric glands ( D , ×100 and F , ×400, hatched areas in D ).

    Journal: Acta Histochemica et Cytochemica

    Article Title: Reactivation of CDX2 in Gastric Cancer as Mark for Gene Silencing Memory

    doi: 10.1267/ahc.15014

    Figure Lengend Snippet: Proper gastric mucosae of fundic gland area ( A , HE, ×100) show negative MUC2 ( B , ×100) and CDX2 ( C , ×100 and E , ×400, hatched areas of C ) expression. MeCP2 expression is, albeit less extensive than surrounding inflammatory cells, positive in the nuclei of proper gastric glands ( D , ×100 and F , ×400, hatched areas in D ).

    Article Snippet: The specimens were then incubated with 2% non-fat dry milk in phosphate-buffered saline (PBS) for 10 min and with primary antibodies against CDX2 (M3636, DAKO, Tokyo, Japan), MUC2 (NBP1-31231, Novus Biologicals, USA, CO) and MeCP2 (D4F3, Cell Signaling Technology, Inc., USA, MA) for 15 min. After three 10-min washes with PBS, the specimens were incubated with rabbit anti-mouse IgG antibody preabsorbed with non-immunized serum.

    Techniques: Expressing

    In the intestinal metaplastic area ( A , HE, ×100), MUC2 ( B , ×100) expression is colocalized with that of CDX2 ( C , ×100 and E , hatched areas of C , ×400). Compared with the non-metaplastic glands (Fig. 1 D and F), and surrounding inflammatory cells, MeCP2 expression is positive in nuclei of the glands in intestinal metaplastic area, but the extent is much less than that in proper gastric mucosa.

    Journal: Acta Histochemica et Cytochemica

    Article Title: Reactivation of CDX2 in Gastric Cancer as Mark for Gene Silencing Memory

    doi: 10.1267/ahc.15014

    Figure Lengend Snippet: In the intestinal metaplastic area ( A , HE, ×100), MUC2 ( B , ×100) expression is colocalized with that of CDX2 ( C , ×100 and E , hatched areas of C , ×400). Compared with the non-metaplastic glands (Fig. 1 D and F), and surrounding inflammatory cells, MeCP2 expression is positive in nuclei of the glands in intestinal metaplastic area, but the extent is much less than that in proper gastric mucosa.

    Article Snippet: The specimens were then incubated with 2% non-fat dry milk in phosphate-buffered saline (PBS) for 10 min and with primary antibodies against CDX2 (M3636, DAKO, Tokyo, Japan), MUC2 (NBP1-31231, Novus Biologicals, USA, CO) and MeCP2 (D4F3, Cell Signaling Technology, Inc., USA, MA) for 15 min. After three 10-min washes with PBS, the specimens were incubated with rabbit anti-mouse IgG antibody preabsorbed with non-immunized serum.

    Techniques: Expressing

    In the area where por elements are seen in a portion of the wel area ( A , ×100), without its target gene, MUC2 expression ( B , ×100), strong CDX2 expression (reactivation, C , ×100, E , ×400, hatched area of C ) is evident in the nuclei of the por elements. MeCP2 expression is almost negative in the por areas ( D , ×100 and F , ×400, hatched areas of D ).

    Journal: Acta Histochemica et Cytochemica

    Article Title: Reactivation of CDX2 in Gastric Cancer as Mark for Gene Silencing Memory

    doi: 10.1267/ahc.15014

    Figure Lengend Snippet: In the area where por elements are seen in a portion of the wel area ( A , ×100), without its target gene, MUC2 expression ( B , ×100), strong CDX2 expression (reactivation, C , ×100, E , ×400, hatched area of C ) is evident in the nuclei of the por elements. MeCP2 expression is almost negative in the por areas ( D , ×100 and F , ×400, hatched areas of D ).

    Article Snippet: The specimens were then incubated with 2% non-fat dry milk in phosphate-buffered saline (PBS) for 10 min and with primary antibodies against CDX2 (M3636, DAKO, Tokyo, Japan), MUC2 (NBP1-31231, Novus Biologicals, USA, CO) and MeCP2 (D4F3, Cell Signaling Technology, Inc., USA, MA) for 15 min. After three 10-min washes with PBS, the specimens were incubated with rabbit anti-mouse IgG antibody preabsorbed with non-immunized serum.

    Techniques: Expressing

    In the superficial wel area ( A , HE, ×100), while MUC2 expression is almost negative ( B , ×100), CDX2 ( C , ×100 and E , ×400, hatched areas of C ) expression is partly but clearly preserved. MeCP2 expression ( D , ×100 and F , ×400, hatched areas in D ) is negative in most of the wel glands.

    Journal: Acta Histochemica et Cytochemica

    Article Title: Reactivation of CDX2 in Gastric Cancer as Mark for Gene Silencing Memory

    doi: 10.1267/ahc.15014

    Figure Lengend Snippet: In the superficial wel area ( A , HE, ×100), while MUC2 expression is almost negative ( B , ×100), CDX2 ( C , ×100 and E , ×400, hatched areas of C ) expression is partly but clearly preserved. MeCP2 expression ( D , ×100 and F , ×400, hatched areas in D ) is negative in most of the wel glands.

    Article Snippet: The specimens were then incubated with 2% non-fat dry milk in phosphate-buffered saline (PBS) for 10 min and with primary antibodies against CDX2 (M3636, DAKO, Tokyo, Japan), MUC2 (NBP1-31231, Novus Biologicals, USA, CO) and MeCP2 (D4F3, Cell Signaling Technology, Inc., USA, MA) for 15 min. After three 10-min washes with PBS, the specimens were incubated with rabbit anti-mouse IgG antibody preabsorbed with non-immunized serum.

    Techniques: Expressing