Fig 1: DSS-induced acute colitis in mice lacking GP2 in gut-associated lymphoid tissue.a VIL1 expression in pancreas, Peyer’s patch (PP), and colon of Vil1-cre-tdTomato mice were shown. b GP2 (red) distribution in Vil1-cre-Gp2flox/flox (GP2IEC) mice were shown. Green color indicates UEA-1 in PP. Representative data of three independent experiments are shown. Scale bars, 100 µm. c Body weight change after induction of acute colitis in GP2IEC and control (WT Gp2flox/flox) mice (n = 7 /group). N.S. indicates not significant (two-tailed unpaired t-test). Data are presented as mean values ± SEM. d Colon length in GP2IEC and control mice were shown (n = 7 /group). N.S. indicates not significant (two-tailed unpaired t-test). Data are presented as mean values ± SEM. e Hematoxylin and eosin staining of colon tissues were shown. Scale bars, 100 µm. Data are representative of three independent experiments. Source data are provided as a Source Data file.
Fig 2: Pancreatic GP2 is important for mucosal protection.a PTF1A-expressing cells (red: tomato) in the pancreas and colon of Ptf1a-creETRM-tdTomato mice. Scale bars, 100 µm. Data are representative of three independent experiments. b Body weight changes after the induction of acute colitis in Ptf1a-creETR ?/+-Gp2flox/flox (Gp2Panc) (n = 15) and control (wild-type; Ptf1a-creETR +/+-Gp2flox/flox) (n = 11) mice, receiving tamoxifen **p < 0.01, *p < 0.05 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. c Changes in colon length and photographs of representative colons of Gp2Panc (n = 15) and control (n = 11) mice. Scale bars, 100 µm. *p = 0.026 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. d Flow cytometric analysis of neutrophils that had infiltrated the colon in Gp2Panc and control mice; representative data are shown. e Hematoxylin and eosin staining of colon at day 8 of DSS treatment in Gp2Panc and control mice. Scale bars: 100 µm. Data are representative of three independent experiments. f Immunohistochemical analysis of colitis colon and luminal contents in Gp2Panc and control mice. Bacteria; EUB338 (red), MUC2 (green), DAPI (blue). Scale bars: upper panel, 100 µm; lower panel, 20 µm. Data are representative of three independent experiments. Source data are provided as a Source Data file.
Fig 3: Recognition of intestinal bacteria by pancreatic GP2.a Luminal contents of colon were stained with GP2 (red) and EUB338 (blue in left bottom and green in right bottom) were shown. Data are representative of three independent experiments. b Fecal unbound GP2 in WT and Gp2–/– mice with or without acute colitis were measured by ELISA (WT, n = 8; Gp2–/–, n = 4). N.S. indicates not significant (two-tailed unpaired t-test). Data are presented as mean values ± SEM. c Percentage of rGP2-bound fecal bacteria isolated from Gp2–/– mice, PBS and rGP2 Intact n = 7, PBS colitis (n = 7), rGP2 colitis (n = 6). *p = 0.017 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. d Representative flow cytometry analysis of fecal bacteria isolated from WT and Gp2–/– mice with or without colitis were shown. e E. coli number of non-selected (rGP2 MACS -) and selected by MACS (rGP2 MACS +) fecal bacteria (n = 9 /group). N.D.: not detected. Data are presented as mean values ± SEM. Source data are provided as a Source Data file.
Fig 4: Upregulation of pancreatic GP2 production in colitis.a Hematoxylin and eosin staining of pancreas in intact or colitis mice are shown. Scale bars, 100 µm. Data are representative of at least three independent experiments. Gene expression of Pnlip in pancreas (b) and of Gp2 in the indicated tissue and cellular components of the gastrointestinal tract (c) in intact mice and mice with acute colitis (n = 4/group, two-tailed unpaired t-test). *p = 0.021. Data are presented as mean values ± SEM. N.S. indicates not significant. d GP2 expression in pancreas. Representative data of four independent experiments are shown. GP2 (red), DAPI (blue). Scale bars, 100 µm. e Concentrations of secretory GP2 in pancreatic juice and intestinal lumen, as measured by enzyme-linked immunosorbent assay (ELISA) (intact, n = 10; colitis, n = 7, two-tailed Mann–Whitney U test). Data are presented as mean values ± SEM. f Fecal GP2 concentration was determined by ELISA and compared between healthy people (n = 5), CD patients (n = 7), and ulcerative colitis patients (n = 9), two-tailed Mann–Whitney U test. Data are presented as mean values ± SEM. Source data are provided as a Source Data file.
Fig 5: Deletion of systemic GP2 results in severe colitis.a Fecal total IgA concentration in WT and Gp2–/– mice, as determined by ELISA (n = 5). N.S. not significant (two-tailed unpaired t-test). Data are presented as mean values ± SEM. b 16S rRNA gene sequencing of fecal bacteria. Phylum levels of bacteria are shown. c qPCR analysis of mucosal bacteria from WT (n = 9) and Gp2–/– (n = 8) mice were examined. *p < 0.05., N.S. indicates not significant (two-tailed unpaired t-test). Data are presented as mean values ± SEM. d Survival ratio and body weight change of WT and Gp2–/– mice with acute colitis (n = 12/group). *p < 0.05, **p < 0.01 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. e Colon length and representative pictures of colon are shown, intact; WT (n = 3), KO (n = 4), and colitis; WT, KO (n = 12). ***p < 0.0001 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. f Hematoxylin and eosin staining of colon at day 8 of DSS treatment are shown. Scale bars: 100 µm. Data are representative of three independent experiments. g Flow cytometry analysis of infiltrated neutrophils is shown. Representative data are shown. h Immunohistochemical analysis of colitis (DSS 2%, day 8) colon with luminal contents of WT and Gp2–/– mice. Bacteria (red), MUC2 (green), DAPI (blue). Scale bars: upper panels, 100 µm; lower panels, 20 µm. Data are representative of three independent experiments. i Serum anti-E. coli IgM, intact (n = 4), and colitis (n = 5), IgA, intact WT (n = 6), KO (n = 10), and colitis WT (n = 15), KO (n = 11), IgG intact WT (n = 12), KO (n = 4), and colitis WT (n = 11), KO (n = 7) in intact and colitis WT and Gp2–/– mice were analyzed by ELISA (Kruskal-Wallis test followed by Mann–Whitney U test). Data are presented as mean values ± SEM. j Body weight changes after the induction of acute TNBS-induced colitis in WT and Gp2–/– mice. ***p < 0.01 (two-tailed unpaired t-test). Data are presented as mean values ± SEM. k Immunohistochemical analysis of colon and luminal contents during TNBS-induced colitis in Gp2Panc and control mice. EUB338 bacteria (red), MUC2 (green), DAPI (blue). Scale bars: upper panels, 100 µm; lower panels, 20 µm. Data are representative of two independent experiments. Source data are provided as a Source Data file.
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