- 38-9200 - Invitrogen Antibodies CLDN15 antibody

Submitted references The dual GLP-1 and GLP-2 receptor agonist dapiglutide promotes barrier function in murine short bowel.

Reiner J, Thiery J, Held J, Berlin P, Skarbaliene J, Vollmar B, Jaster R, Eriksson PO, Lamprecht G, Witte M
Annals of the New York Academy of Sciences 2022 Aug;1514(1):132-141

Nanoscale segregation of channel and barrier claudins enables paracellular ion flux.

Gonschior H, Schmied C, Van der Veen RE, Eichhorst J, Himmerkus N, Piontek J, Günzel D, Bleich M, Furuse M, Haucke V, Lehmann M
Nature communications 2022 Aug 25;13(1):4985

Inactivation of paracellular cation-selective claudin-2 channels attenuates immune-mediated experimental colitis in mice.

Raju P, Shashikanth N, Tsai PY, Pongkorpsakol P, Chanez-Paredes S, Steinhagen PR, Kuo WT, Singh G, Tsukita S, Turner JR
The Journal of clinical investigation 2020 Oct 1;130(10):5197-5208

Differential regulation of claudin-2 and claudin-15 expression in children and adults with malabsorptive disease.

Ong MLDM, Yeruva S, Sailer A, Nilsen SP, Turner JR
Laboratory investigation; a journal of technical methods and pathology 2020 Mar;100(3):483-490

EspF is crucial for Citrobacter rodentium-induced tight junction disruption and lethality in immunocompromised animals.

Xia X, Liu Y, Hodgson A, Xu D, Guo W, Yu H, She W, Zhou C, Lan L, Fu K, Vallance BA, Wan F
PLoS pathogens 2019 Jun;15(6):e1007898

Vitamin D Receptor Deletion Leads to the Destruction of Tight and Adherens Junctions in Lungs.

Chen H, Lu R, Zhang YG, Sun J
Tissue barriers 2018;6(4):1-13

IL-22 Upregulates Epithelial Claudin-2 to Drive Diarrhea and Enteric Pathogen Clearance.

Tsai PY, Zhang B, He WQ, Zha JM, Odenwald MA, Singh G, Tamura A, Shen L, Sailer A, Yeruva S, Kuo WT, Fu YX, Tsukita S, Turner JR
Cell host & microbe 2017 Jun 14;21(6):671-681.e4

Obesity-induces Organ and Tissue Specific Tight Junction Restructuring and Barrier Deregulation by Claudin Switching.

Ahmad R, Rah B, Bastola D, Dhawan P, Singh AB
Scientific reports 2017 Jul 11;7(1):5125

Disrupted tight junctions in the small intestine of cystic fibrosis mice.

De Lisle RC
Cell and tissue research 2014 Jan;355(1):131-42

EpCAM contributes to formation of functional tight junction in the intestinal epithelium by recruiting claudin proteins.

Lei Z, Maeda T, Tamura A, Nakamura T, Yamazaki Y, Shiratori H, Yashiro K, Tsukita S, Hamada H
Developmental biology 2012 Nov 15;371(2):136-45

Dynamics of claudins expression in colitis and colitis-associated cancer in rat.

Arimura Y, Nagaishi K, Hosokawa M
Methods in molecular biology (Clifton, N.J.) 2011;762:409-25

Loss of hepatocyte-nuclear-factor-4alpha affects colonic ion transport and causes chronic inflammation resembling inflammatory bowel disease in mice.

Darsigny M, Babeu JP, Dupuis AA, Furth EE, Seidman EG, Lévy E, Verdu EF, Gendron FP, Boudreau F
PloS one 2009 Oct 29;4(10):e7609

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Supportive validation

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Experimental details: Immunofluorescent co-labeling (yellow) of claudin-15 (red) and ZO-1 (green) in mouse ileum tissue using Rb anti-Claudin-15 (Product # 38-9200). Image courtesy of Jennifer Holmes and Dr. James Anderson, University of North Carolina at Chapel Hill, NC.

Supportive validation

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Experimental details: Immunofluorescent staining of (A) Anti-Claudin 15; (B) Claudin 15 (red), ZO-1 (Green) co localization (yellow); (C) Anti-Claudin 15; (D) Claudin 15 (red), ZO-1 (green) -- C and D (blocked by fusion protein) using Rb anti-Claudin-15 (Product # 38-9200).

Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Claudin-15 and ion transport are modulated in Hnf4alpha colon mutants. (A) qRT-PCR of claudin-4, -8 and -15 at various time points in mutants (Open bars) relative to controls (Black bars). Expression levels are shown as mean values (+- SEM) relative to controls at each time point (3-14 mice per group) and were normalized with the Tbp housekeeping gene. (B) Immunoblotting confirmed modulated protein levels of claudin-4, -8 and -15 at 12 months in mutants (n = 3, open bars) and relative controls (n = 3, black bars) normalized to stable expression of E-Cadherin (Cdh1). (C) Quantification of band intensities of each claudin (means +- SEM; n = 3 or 4 for each group) relative to Cdh1. (D) Indirect immunofluorescence (FITC) of claudin-4, -8 and -15 in 3 month-old animals in proximal colon (representative of 4 pairs). Nuclei were counterstained with DAPI (blue) and merged with FITC (green). Scale bars, 50 um. (E) Claudin-15 expression is lost in the upper half of the colon epithelium in mutants (Open bar) relative to controls (Black bar). (F) Colonic permeability to 51 Cr-EDTA measured in colon tissues from 2 month-old mice (5 mice per group). (G) Baseline short circuit current (Isc) measured in colon tissues from 2 month-old mice (5 mice per group). Represented as mean values +- SEM, * P

Supportive validation

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Experimental details: Figure 6 The claudin-15 gene is a direct target of Hnf4alpha. (A) EMSA of 4 Hnf4alpha binding sites found in the 1 kb mouse claudin-15 promoter performed with 293T cells transfected with pBabe Hnf4alpha1 vector and 293T nuclear extracts as negative controls. Shifts (white arrow head) are compared to previously published APOC3 and mutated APOC3 (APOC3m) for control binding site. NS is non-specific binding (grey arrow head). Supershifts (black arrow head) were done with Hnf4alpha antibody (sc-6556). (B) ChIP assays of the 4 putative sites performed on epithelial cells from adult mouse colon. (C) qRT-PCR of claudin-15 on IEC6 cells infected with pBabe Hnf4alpha as compared to control vector pBabe (n = 4). (D) Western blot of 3 stable T84 cell populations expressing mouse claudin-15. (E) Conductance measured for three consecutive days after the reach of confluence on three populations of T84 cells expressing mouse claudin-15 or control plenti eGFP vector. Represented as mean values+-SEM, *** P

Supportive validation

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Experimental details: 3 FIGURE Dapiglutide promotes cation selectivity via expansion of cation-selective claudins in the epithelium. (A) Biionic dilution potentials were measured in the Ussing chamber by replacing 50% NaCl in the basal solution with counter-cation chloride in increasing size, n = 5-6 per group. Mean +- SEM, * p < 0.05 versus ICR + vehicle, SS p < 0.05 for ICR + vehicle versus ICR + dapiglutide. (B) Upper panel: representative western blot of claudin-2 protein of jejunum tissue. Lower panel: corresponding densitometric quantification, n = 4-5 per group, mean +- SEM, * p < 0.05, ** p < 0.01. (C) Upper panel: representative western blot of claudin-15 protein of jejunum tissue. Lower panel: corresponding densitometric quantification, n = 4-5 per group, mean +- SEM. (D) Immunofluorescence images of jejunum sections: claudin staining in green, F-actin in red, and nuclei in blue. Scale bar: 50 mum. White arrows indicate the extension of the indicated claudin along the crypt-villus axis

Supportive validation

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Main image
Experimental details: Claudins form characteristic copolymers including segregation of channel-forming claudins from Cldn3. a Scheme is illustrating predicted claudin-claudin organization patterns. Two claudins (yellow and magenta) that are compatible with each other tightly colocalize in the TJ (pink). Incompatible claudins separate into different strands or even larger claudin-specific parts. This separation might facilitate permeability of specific ions (cyan) over the TJ meshwork. b Representative STED images of TJ-like meshwork of the observed five organization patterns formed by the indicated co-expressed claudins (SNAP-tagged (yellow; BG-Atto590) and YFP-tagged (magenta; alpha-GFP-NB-Atto647N)) in fixed COS-7 cells are shown. Tight colocalization: intermixing of mesh-forming claudins, integration of non-mesh-forming with a mesh-forming claudin or induction (de novo mesh-forming of co-expressed non-mesh-forming claudins). Separated claudins: segregation and exclusion. c Pearson correlation analysis of Cldn3 co-expressed with barrier-forming (gray) or channel-forming (magenta) claudins. Co-expression of Cldn3 with Cldn3 (yellow) served as positive control. Data represent the mean +- SD. Every n represents the Pearson of one TJ-like meshwork. n (Cldn3 + Cldn3) = 55; n (Cldn3 + Cldn1) = 54; n (Cldn3 + Cldn5) = 20; n (Cldn3 + Cldn6) = 20; n (Cldn3 + Cldn7) = 17; n (Cldn3 + Cldn9) = 15; n (Cldn3 + Cldn14) = 19; n (Cldn3 + Cldn19b) = 25; n (Cldn3 + Cldn2) = 36; n (Cldn3 + Cldn10a) = 30; n (Cldn3 +

Supportive validation

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Main image
Experimental details: Segregation enables paracellular ion flux over the TJ meshwork. a Representative confocal images of MDCKII QKO cells stably expressing (sT) FLAG-Cldn2, FLAG-Cldn10a, and FLAG-Cldn2+FLAG-Cldn10a. Single claudin expressing cells were immunostained with anti-Cldn2 (magenta; 2 nd -Atto647N) or anti-Cldn10 (magenta; 2 nd -Atto647N) and anti-ZO-1 (yellow; 2 nd -AF594) antibody. Double claudin expressing cells were immunostained with anti-Cldn2 (magenta; 2 nd -Atto647N) and anti-Cldn10 (yellow; 2 nd -AF594). The white rectangle indicates the region of interest for the magnification in ( b ). b Magnification of FLAG-Cldn2+FLAG-Cldn10a expressing cells from ( a ). White arrows point out differently sized TJ parts that contain only Cldn10a. c Representative confocal images of MDCKII QKO sT FLAG-Cldn3, FLAG-Cldn15 and FLAG-Cldn3+FLAG-Cldn15. Single and double claudin expressing cells were stained with anti-Cldn3 (magenta; 2 nd -Atto647N) or anti-Cldn15 (magenta; 2 nd -Atto647N) and anti-ZO-1 (yellow; 2 nd -AF594) antibody. d Summary of the TER (ohm*cm 2 ), fluorescein flux (10 -6 cm/s) and PNa/PCl ratio from the measured cell lines: MDCKII, MDCKII QKO and MDCKII QKO sT FLAG-tagged Cldn2, Cldn3, Cldn10a, Cldn15 and MDCKII QKO sT FLAG-tagged Cldn2 + Cldn10a and FLAG-tagged Cldn3 + Cldn15. Data represent the mean +- SD from 3 to 5 independent experiments. Single data points are shown in Supplementary Fig. 12 . e Absolute permeability for sodium (PNa; magenta bars) and chloride (PCl; cyan b

38-9200

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