antibody occludin Search Results


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  • 99
    Thermo Fisher occludin
    Gelatinase activity in BALF and expression of claudin-4 and <t>occludin</t> in the three experimental groups. ( a ) Bands corresponding to MMP-2 (63 kDa) and MMP-9 (80 kDa) are highlighted. ( b ) Quantitative data for MMP-9 are presented as mean ± SD. ( c ) Western blots of tissue samples show the difference between groups in claudin-4 abundance. ( d ) Quantitative data for claudin-4 in each group were normalized to β-actin. ( e ) Western blots of tissue samples show difference between groups in occludin abundance. ( f ) Quantitative data for occludin for each group were normalized using β-actin. * p
    Occludin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3688 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc occludin
    Downregulation of tight junction proteins following peripheral nerve injury. Microvessels isolated from the spinal cord were positive for endothelium markers, VWF and Glut-1 ( A ). The expression of tight junction-associated protein ZO-1 ( B ), <t>occludin-1</t> ( C ), and caveolin-1, a major player in the formation of caveolae ( D ), was significantly lower in microvessels of animals day 3 after nerve lesion than controls (naive and sham). Quantitative assessment of protein levels is illustrated in E . Three separate experiments in which each treatment group consists of pooled microvessels from two animals were included. Values are presented as means ± SEM. * p
    Occludin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 168 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Cell Signaling Technology Inc occludin antibodies
    QYT relieves the reduced expression of junction proteins in vascular endothelial cells. (A) The immunofluorescence confocal pictures showing claudin-5. Claudin-5 (red) localized at peripheral of the endothelial cells which were marked by VWF (green). Blue color shows nuclei. The rectangle region in each picture numbered 1 is magnified and shown below as pictures numbered 2, correspondingly. Bars = 50 μm in (a1-f1) , Bars = 7.5 μm in (a2-f2) . (B) Representative western blots of claudin-5, <t>occludin,</t> JAM-1, ZO-1 and VE-cadherin in different groups. Shown on the right side is the quantification of claudin-5 (C) , occludin (E) , JAM-1 (F) , ZO-1 (G) , VE-cadherin (H) , respectively. (D) The quantitative analysis of claudin-5 fluorescent intensity. All the quantifications were performed based on the data of four independent experiments and normalized to GAPDH, respectively. Values are the mean ± SEM. * p
    Occludin Antibodies, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human occludin antibody
    Analysis of barrier formation in endothelial monolayer. (A) Expression of endothelial junctional molecules CD31, CD144 and ZO-1 (from left to right) and actin (far right) in hCMVEC monolayer 48 h after seeding. (B) The permeability of hCMVEC monolayer to FITC-dextran 48 h after seeding (mean ± SEM, n = 4). (C) Time course analysis of endothelial tight junction molecule <t>occludin</t> expression at 24, 48 and 72 h post seeding of hCMVECs using Western blot using a mouse monoclonal anti-occludin (left) and rabbit polyclonal anti-occludin (right). (D) Real time measurement of electrical resistance across the endothelial monolayer as an indicator of endothelial barrier integrity. The barrier resistance (Rb) and the basolateral adhesion (Alpha) due to focal adhesions can be modelled using the ECIS Z theta software when the experiment is run using multi-frequency mode (250 to 64000 Hz). The modelling reveals that the basolateral adhesion occurs very fast and junction formation begins around 10 hours after seeding. Overall barrier resistance is the summation of both Rb and alpha, but this software function can determine whether barrier changes are predominantly due to the Rb. Data are mean ± SEM (n = 4).
    Human Occludin Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam occludin
    Tight Junction Protein, <t>Occludin,</t> Expression in the Cerebral Vasculature of C57BL/6 Mice is not Altered through Exposure to Mixed Vehicle Emissions Double immunofluorescence of occludin (red) and vWF (green) in cerebral microvessels from 4 mo old C57BL/6 mice on a low-fat (LF; A–C) or high-fat diet (HF; G–I) exposed filtered air (FA) or on a LF (D–F) or HF diet (J–L) and to exposed MVE at 100 µg/m 3 PM of mixed gasoline and diesel engine emissions for 6hr/d, 7 d/wk, for 30 d (n=3–4 per exposure group; 4 sections each, 3–5 vessels per section). Right panels (yellow; C, F, I, L) are merged figures of left (red; A, D, G, J) and center (green, E, H, K) panels for occludin and vWF. Blue fluorescence is Hoechst stained nuclei. Scale bar = 100 µm. Results shown are mean ± SEM.
    Occludin, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1307 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology occludin antibody
    Metformin attenuates Staphylococcus aureus induced cleavage of <t>occludin</t> at 60 kD and alters cleavage fragments at 44 kD. (A) Occludin bands were normalized to β‐actin and to their respective controls,* P
    Occludin Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Proteintech occludin
    FICZ prevents DSS-induced disruption of TJ. (A) Expressions of TJ proteins (ZO-1, <t>Occludin</t> and Claudin-1) and CYP1A1 in the colonic mucosa were evaluated by western blotting. (B) Immunohistochemical analysis of TJ proteins in the colonic mucosa. Magnification: ×400.
    Occludin, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 253 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals occludin
    Overall working model HHcy induced oxidative and ER stress conditions in the eyes of CBS +/− mice indicating the presence of ROS through the upregulation of BiP, PERK and downregulation of TJPs such as <t>occludin</t> respectively. These metabolic changes are generally accompanied by the excess production of glutamate. Such physiological alterations can contribute to the downregulation TJPs making blood vessels permeable which can lead to the loss of ocular homeostasis followed by vision impairment. GYY4137 treatment could potentially mitigate these harmful effects.
    Occludin, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novus Biologicals occludin antibody 2b3
    Overall working model HHcy induced oxidative and ER stress conditions in the eyes of CBS +/− mice indicating the presence of ROS through the upregulation of BiP, PERK and downregulation of TJPs such as <t>occludin</t> respectively. These metabolic changes are generally accompanied by the excess production of glutamate. Such physiological alterations can contribute to the downregulation TJPs making blood vessels permeable which can lead to the loss of ocular homeostasis followed by vision impairment. GYY4137 treatment could potentially mitigate these harmful effects.
    Occludin Antibody 2b3, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Selleck Chemicals occludin
    Effects of resveratrol on H 2 O 2 -induced cytotoxicity in Nrf2-knockdown IPEC-J2 cells. The Nrf2-knockdown (shNRF2-IPEC-J2) or control (SC-IPEC-J2) cells were pretreated with or without 50 μ M RSV for 6 hours and then cocultured with 500 μ M H 2 O 2 for 4 hours. (a) Cell viability was measured using the CCK-8 assay. Results are presented as the percentage of cell viability compared with the control (0 μ M). (b, c) The relative expression of SOD-1, CAT, claudin-1, <t>occludin,</t> and ZO-1 was detected by qRT-PCR. Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (d) Apoptotic cells were analyzed by flow cytometry using Annexin V-PI double staining. (e) Protein levels of Nrf2, Akt, p-Nrf2, and p-Akt were detected by Western blot with β -actin as the loading control. Values are the mean ± SE. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P
    Occludin, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Biorbyt occludin antibody
    Immunohistochemical expression of <t>occludin</t> in the colonic mucosa of patients with (a) non-CKD, (b) advanced CKD, and (c) HD with hypotension. The total of epithelial cells lining villi exhibited positive immunostaining for occludin in healthy controls (a). In patients with advanced CKD, occludin expression is reduced in numerous epithelial cells (b). In HD patients with IDH, there was profound loss of occludin expression by most epithelial cells lining villi (blue arrows) (c).
    Occludin Antibody, supplied by Biorbyt, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher occludin antibodies
    Raft localization of <t>occludin.</t> Caco-2 cells were untreated (A), treated with CPE (2.5 μg/ml) for 60 min (B), or treated for 60 min with 10 mM MβCD (C), followed by cold Triton X-100 extraction and sucrose density gradient centrifugation.
    Occludin Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson occludin
    Nectin-2, but not <t>occludin,</t> is expressed in RV, comma-shaped bodies, and S-shaped bodies. Five-micrometer sections of e18 mouse kidney were stained with monoclonal anti-nectin-2 antibody ( A ′, A ′′′, and B – E ) and polyclonal anti-occludin antibody ( A , A ′′′, and B – E ), Hoechst 33342 ( A ′′, A ′′′, and B – D ), and lotus lectin (LTL; blue staining, E ). UB and RV are labeled using arrows. Open arrowheads demonstrate punctate nectin-2 and occludin staining in UB ( A – A ′′′). In B at high magnification, yellow indicates colocalization of nectin-2 and occludin, and green and red indicate areas where nectin-2 and occludin did not colocalize (open arrowheads and filled arrowheads, respectively). Asterisks demonstrate RV, comma-shaped body, and S-shaped body where nectin-2 is expressed, but occludin is not ( A – A ′′′, C , and D ). Scale bars = 10 μm.
    Occludin, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 183 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Bioss occludin
    KO of LRRC75A-AS1 enhanced TJ of MAC-T cells. (A) Gene expressions of Claudin-1, <t>Occludin</t> and ZO-1 were analyzed by RT-qPCR. (B) Claudin-1, Occludin and ZO-1 expression in KO-Control and KO-LRRC75A-AS1 cells were analyzed by western blot. GAPDH was used as an internal control. (C) The relative quantitative analysis of western blot result was performed. *P
    Occludin, supplied by Bioss, used in various techniques. Bioz Stars score: 90/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bioss occludin polyclonal antibody
    KO of LRRC75A-AS1 enhanced TJ of MAC-T cells. (A) Gene expressions of Claudin-1, <t>Occludin</t> and ZO-1 were analyzed by RT-qPCR. (B) Claudin-1, Occludin and ZO-1 expression in KO-Control and KO-LRRC75A-AS1 cells were analyzed by western blot. GAPDH was used as an internal control. (C) The relative quantitative analysis of western blot result was performed. *P
    Occludin Polyclonal Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Gelatinase activity in BALF and expression of claudin-4 and occludin in the three experimental groups. ( a ) Bands corresponding to MMP-2 (63 kDa) and MMP-9 (80 kDa) are highlighted. ( b ) Quantitative data for MMP-9 are presented as mean ± SD. ( c ) Western blots of tissue samples show the difference between groups in claudin-4 abundance. ( d ) Quantitative data for claudin-4 in each group were normalized to β-actin. ( e ) Western blots of tissue samples show difference between groups in occludin abundance. ( f ) Quantitative data for occludin for each group were normalized using β-actin. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Adaptive Support Ventilation Attenuates Ventilator Induced Lung Injury: Human and Animal Study

    doi: 10.3390/ijms20235848

    Figure Lengend Snippet: Gelatinase activity in BALF and expression of claudin-4 and occludin in the three experimental groups. ( a ) Bands corresponding to MMP-2 (63 kDa) and MMP-9 (80 kDa) are highlighted. ( b ) Quantitative data for MMP-9 are presented as mean ± SD. ( c ) Western blots of tissue samples show the difference between groups in claudin-4 abundance. ( d ) Quantitative data for claudin-4 in each group were normalized to β-actin. ( e ) Western blots of tissue samples show difference between groups in occludin abundance. ( f ) Quantitative data for occludin for each group were normalized using β-actin. * p

    Article Snippet: The membranes were then incubated with antibodies to claudin-4 (1:100 dilution), occludin (1:100 dilution; both from Invitrogen, Carlsbad, CA, USA), and β-actin (1:10,000 dilution; Sigma-Aldrich, St. Louis, MO, USA) overnight at 4 °C with mild shaking, followed by incubation of membranes with horseradish peroxidase-conjugated anti-rabbit (1:20,000 dilution) and anti-mouse (1:50,000 dilution; both from Jackson Laboratory, Bar Harbor, ME, USA), respectively, for 1 h at RT with gentle shaking.

    Techniques: Activity Assay, Expressing, Western Blot

    Downregulation of tight junction proteins following peripheral nerve injury. Microvessels isolated from the spinal cord were positive for endothelium markers, VWF and Glut-1 ( A ). The expression of tight junction-associated protein ZO-1 ( B ), occludin-1 ( C ), and caveolin-1, a major player in the formation of caveolae ( D ), was significantly lower in microvessels of animals day 3 after nerve lesion than controls (naive and sham). Quantitative assessment of protein levels is illustrated in E . Three separate experiments in which each treatment group consists of pooled microvessels from two animals were included. Values are presented as means ± SEM. * p

    Journal: The Journal of Neuroscience

    Article Title: Peripheral Nerve Injury Alters Blood–Spinal Cord Barrier Functional and Molecular Integrity through a Selective Inflammatory Pathway

    doi: 10.1523/JNEUROSCI.1642-11.2011

    Figure Lengend Snippet: Downregulation of tight junction proteins following peripheral nerve injury. Microvessels isolated from the spinal cord were positive for endothelium markers, VWF and Glut-1 ( A ). The expression of tight junction-associated protein ZO-1 ( B ), occludin-1 ( C ), and caveolin-1, a major player in the formation of caveolae ( D ), was significantly lower in microvessels of animals day 3 after nerve lesion than controls (naive and sham). Quantitative assessment of protein levels is illustrated in E . Three separate experiments in which each treatment group consists of pooled microvessels from two animals were included. Values are presented as means ± SEM. * p

    Article Snippet: After the transfer, blots were incubated overnight at 4°C with the following antibodies: zonula occludens-1 (ZO-1) (1:200; Cell Signaling), occludin (1:100; Cell Signaling), caveolin-1 (1:250; Cell Signaling), TGF-β receptor I (TGF-β-RI) (1:200; Santa Cruz), and p-Smad2/3 (1:500; Santa Cruz).

    Techniques: Isolation, Expressing

    TGF-β1 alters tight junction protein levels in spinal cord microvessels. Intrathecal administration of TGF-β1 (2.5 μg for day 0 to day 3) successfully prevents the decrease in tight junction protein levels ZO-1 ( A , D ) and occludin ( B , D ) observed after peripheral nerve injury. Levels of caveolae structural component caveolin-1, however, remained unchanged ( C , D ). The receptor for TGF-β1 (TGF-β-RI) is found in isolated spinal cord endothelial cells, confirmed by the colocalization with vascular marker CD31. DAPI staining (blue) was used for cellular identification ( E ). The protein levels of the TGF-β-RI remain unchanged after peripheral nerve injury, with or without TGF-β1 infusion ( F ). TGF-β1 treatment induced the phosphorylation of signaling pathway proteins Smad2/3 (pSmad2/3) in spinal cord microvessels ( G ). Three separate experiments in which each treatment group consists of pooled microvessels from two animals were included. Values are presented as means ± SEM. * p

    Journal: The Journal of Neuroscience

    Article Title: Peripheral Nerve Injury Alters Blood–Spinal Cord Barrier Functional and Molecular Integrity through a Selective Inflammatory Pathway

    doi: 10.1523/JNEUROSCI.1642-11.2011

    Figure Lengend Snippet: TGF-β1 alters tight junction protein levels in spinal cord microvessels. Intrathecal administration of TGF-β1 (2.5 μg for day 0 to day 3) successfully prevents the decrease in tight junction protein levels ZO-1 ( A , D ) and occludin ( B , D ) observed after peripheral nerve injury. Levels of caveolae structural component caveolin-1, however, remained unchanged ( C , D ). The receptor for TGF-β1 (TGF-β-RI) is found in isolated spinal cord endothelial cells, confirmed by the colocalization with vascular marker CD31. DAPI staining (blue) was used for cellular identification ( E ). The protein levels of the TGF-β-RI remain unchanged after peripheral nerve injury, with or without TGF-β1 infusion ( F ). TGF-β1 treatment induced the phosphorylation of signaling pathway proteins Smad2/3 (pSmad2/3) in spinal cord microvessels ( G ). Three separate experiments in which each treatment group consists of pooled microvessels from two animals were included. Values are presented as means ± SEM. * p

    Article Snippet: After the transfer, blots were incubated overnight at 4°C with the following antibodies: zonula occludens-1 (ZO-1) (1:200; Cell Signaling), occludin (1:100; Cell Signaling), caveolin-1 (1:250; Cell Signaling), TGF-β receptor I (TGF-β-RI) (1:200; Santa Cruz), and p-Smad2/3 (1:500; Santa Cruz).

    Techniques: Isolation, Marker, Staining

    QYT relieves the reduced expression of junction proteins in vascular endothelial cells. (A) The immunofluorescence confocal pictures showing claudin-5. Claudin-5 (red) localized at peripheral of the endothelial cells which were marked by VWF (green). Blue color shows nuclei. The rectangle region in each picture numbered 1 is magnified and shown below as pictures numbered 2, correspondingly. Bars = 50 μm in (a1-f1) , Bars = 7.5 μm in (a2-f2) . (B) Representative western blots of claudin-5, occludin, JAM-1, ZO-1 and VE-cadherin in different groups. Shown on the right side is the quantification of claudin-5 (C) , occludin (E) , JAM-1 (F) , ZO-1 (G) , VE-cadherin (H) , respectively. (D) The quantitative analysis of claudin-5 fluorescent intensity. All the quantifications were performed based on the data of four independent experiments and normalized to GAPDH, respectively. Values are the mean ± SEM. * p

    Journal: Frontiers in Physiology

    Article Title: Post-treatment With Qing-Ying-Tang, a Compound Chinese Medicine Relives Lipopolysaccharide-Induced Cerebral Microcirculation Disturbance in Mice

    doi: 10.3389/fphys.2019.01320

    Figure Lengend Snippet: QYT relieves the reduced expression of junction proteins in vascular endothelial cells. (A) The immunofluorescence confocal pictures showing claudin-5. Claudin-5 (red) localized at peripheral of the endothelial cells which were marked by VWF (green). Blue color shows nuclei. The rectangle region in each picture numbered 1 is magnified and shown below as pictures numbered 2, correspondingly. Bars = 50 μm in (a1-f1) , Bars = 7.5 μm in (a2-f2) . (B) Representative western blots of claudin-5, occludin, JAM-1, ZO-1 and VE-cadherin in different groups. Shown on the right side is the quantification of claudin-5 (C) , occludin (E) , JAM-1 (F) , ZO-1 (G) , VE-cadherin (H) , respectively. (D) The quantitative analysis of claudin-5 fluorescent intensity. All the quantifications were performed based on the data of four independent experiments and normalized to GAPDH, respectively. Values are the mean ± SEM. * p

    Article Snippet: Antibodies against occludin, JAM-1, ZO-1, cav-1, phosphor-cav-1, VE-cadherin, and GAPDH were obtained from Cell Signaling Technology (Beverly, MA, United States).

    Techniques: Expressing, Immunofluorescence, Western Blot

    TJ destruction owing to high glucose in RF/6A cells required RhoA/ROCK1 signaling pathway. A: Western blotting (up) and quantification (down) of occludin profiles in five cell treatment groups: NG, HG, high glucose with ROCK1-siRNA (HG + ROCK1-siRNA),

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Involvement of RhoA/ROCK1 signaling pathway in hyperglycemia-induced microvascular endothelial dysfunction in diabetic retinopathy

    doi:

    Figure Lengend Snippet: TJ destruction owing to high glucose in RF/6A cells required RhoA/ROCK1 signaling pathway. A: Western blotting (up) and quantification (down) of occludin profiles in five cell treatment groups: NG, HG, high glucose with ROCK1-siRNA (HG + ROCK1-siRNA),

    Article Snippet: Primary antibodies against occludin, claudin-5 and ZO-1 were purchased from cell signaling technology (Danvers, MA, USA), anti-RhoA and anti-ROCK1, anti-p-MYPT1 (Thr853) and anti-MYPT1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Western Blot

    Effects of increasing concentrations of rifaximin (0.1, 1.0, and 10 μM) alone and rifaximin plus ketoconazole (10 μM) against TcdA (30 ng/ml) in Caco-2 cells: (A) 24-h time course TEER changes ( n = 4); (B) MTT cell viability absorbance at 24 h ( n = 5); (C) Immunoreactive bands corresponding to Bax, ZO-1, and occludin expression at 24 h following the TcdA challenge; (D) Relative densitometric analysis of immunoreactive bands (arbitrary units normalized against the expression of the housekeeping GAPDH protein; n = 5), and (E) Immunofluorescent staining showing the effects of TcdA challenge on ZO-1 and occludin co-expression at 24 h. Nuclei were also investigated using DAPI staining (Scale bar = 25 μm). Results are expressed as mean ± SEM of experiments performed in triplicate. ∗∗∗ p

    Journal: Frontiers in Pharmacology

    Article Title: Rifaximin Improves Clostridium difficile Toxin A-Induced Toxicity in Caco-2 Cells by the PXR-Dependent TLR4/MyD88/NF-κB Pathway

    doi: 10.3389/fphar.2016.00120

    Figure Lengend Snippet: Effects of increasing concentrations of rifaximin (0.1, 1.0, and 10 μM) alone and rifaximin plus ketoconazole (10 μM) against TcdA (30 ng/ml) in Caco-2 cells: (A) 24-h time course TEER changes ( n = 4); (B) MTT cell viability absorbance at 24 h ( n = 5); (C) Immunoreactive bands corresponding to Bax, ZO-1, and occludin expression at 24 h following the TcdA challenge; (D) Relative densitometric analysis of immunoreactive bands (arbitrary units normalized against the expression of the housekeeping GAPDH protein; n = 5), and (E) Immunofluorescent staining showing the effects of TcdA challenge on ZO-1 and occludin co-expression at 24 h. Nuclei were also investigated using DAPI staining (Scale bar = 25 μm). Results are expressed as mean ± SEM of experiments performed in triplicate. ∗∗∗ p

    Article Snippet: Rabbit anti-zona occludens-1 (ZO-1), anti-occludin and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibodies were procured from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: MTT Assay, Expressing, Staining

    NAC attenuates PM-induced tight junction disruption. BEAS-2B cells were pretreated with or without NAC (5 mM) for 1 h before PM (200 µg/mL). Twenty-four hours later, cells were infected with P. aeruginosa (MOI 10) for an additional 2 hours. (A) Immunofluorescence of ZO-1, occludin and claudin-1 (magnification 200); (B) the protein expression of occludin and claudin-1 were detected by western blot; (C,D) the statistical graph of occludin and claudin-1 protein expression; (E) the TEER were measured in an air-liquid model. Data are expressed as mean ± SD, from three separate experiments. **, P

    Journal: Journal of Thoracic Disease

    Article Title: Particulate matter disrupts airway epithelial barrier via oxidative stress to promote Pseudomonas aeruginosa infection

    doi: 10.21037/jtd.2019.05.77

    Figure Lengend Snippet: NAC attenuates PM-induced tight junction disruption. BEAS-2B cells were pretreated with or without NAC (5 mM) for 1 h before PM (200 µg/mL). Twenty-four hours later, cells were infected with P. aeruginosa (MOI 10) for an additional 2 hours. (A) Immunofluorescence of ZO-1, occludin and claudin-1 (magnification 200); (B) the protein expression of occludin and claudin-1 were detected by western blot; (C,D) the statistical graph of occludin and claudin-1 protein expression; (E) the TEER were measured in an air-liquid model. Data are expressed as mean ± SD, from three separate experiments. **, P

    Article Snippet: BEAS-2B cells and lung tissues of mice from each group was obtained and electrophoresed, transferred to the Poly vinylidene fluoride (PVDF) membranes, and then incubated with anti-occludin antibody, anti-claudin-1 antibody, and anti-GADPH antibody (CST, Boston, MA, USA) overnight at 4 °C.

    Techniques: Infection, Immunofluorescence, Expressing, Western Blot

    PM increases P. aeruginosa -infected lung injury, lung bacteria burden and blood bacterial dissemination accompanying by the degradation of the tight junction in vivo. (A) Histological changes of the four group: Control (PBS) and PM (0.5, 2, or 4 mg/kg) (magnification 200); (B) mean lung injury score for each group; (C,D) the protein expression of occluding and claudin-1in mice were detected by western blot. (E) the CFUs in the lungs of mice in the control and PM exposed groups (0.5, 2, or 4 mg/kg) 24 hours post- P. aeruginosa instillation; (F) the CFUs in the blood of mice in the control and PM exposed groups 24 hours post- P. aeruginosa instillation. A total of 5–6 mice were examined per group; (G,H) the correlation analysis of protein abundance of occludin and claudin-1 with the lung bacteria burden. Data are expressed as mean ± SD. *, P

    Journal: Journal of Thoracic Disease

    Article Title: Particulate matter disrupts airway epithelial barrier via oxidative stress to promote Pseudomonas aeruginosa infection

    doi: 10.21037/jtd.2019.05.77

    Figure Lengend Snippet: PM increases P. aeruginosa -infected lung injury, lung bacteria burden and blood bacterial dissemination accompanying by the degradation of the tight junction in vivo. (A) Histological changes of the four group: Control (PBS) and PM (0.5, 2, or 4 mg/kg) (magnification 200); (B) mean lung injury score for each group; (C,D) the protein expression of occluding and claudin-1in mice were detected by western blot. (E) the CFUs in the lungs of mice in the control and PM exposed groups (0.5, 2, or 4 mg/kg) 24 hours post- P. aeruginosa instillation; (F) the CFUs in the blood of mice in the control and PM exposed groups 24 hours post- P. aeruginosa instillation. A total of 5–6 mice were examined per group; (G,H) the correlation analysis of protein abundance of occludin and claudin-1 with the lung bacteria burden. Data are expressed as mean ± SD. *, P

    Article Snippet: BEAS-2B cells and lung tissues of mice from each group was obtained and electrophoresed, transferred to the Poly vinylidene fluoride (PVDF) membranes, and then incubated with anti-occludin antibody, anti-claudin-1 antibody, and anti-GADPH antibody (CST, Boston, MA, USA) overnight at 4 °C.

    Techniques: Infection, In Vivo, Expressing, Mouse Assay, Western Blot

    Analysis of barrier formation in endothelial monolayer. (A) Expression of endothelial junctional molecules CD31, CD144 and ZO-1 (from left to right) and actin (far right) in hCMVEC monolayer 48 h after seeding. (B) The permeability of hCMVEC monolayer to FITC-dextran 48 h after seeding (mean ± SEM, n = 4). (C) Time course analysis of endothelial tight junction molecule occludin expression at 24, 48 and 72 h post seeding of hCMVECs using Western blot using a mouse monoclonal anti-occludin (left) and rabbit polyclonal anti-occludin (right). (D) Real time measurement of electrical resistance across the endothelial monolayer as an indicator of endothelial barrier integrity. The barrier resistance (Rb) and the basolateral adhesion (Alpha) due to focal adhesions can be modelled using the ECIS Z theta software when the experiment is run using multi-frequency mode (250 to 64000 Hz). The modelling reveals that the basolateral adhesion occurs very fast and junction formation begins around 10 hours after seeding. Overall barrier resistance is the summation of both Rb and alpha, but this software function can determine whether barrier changes are predominantly due to the Rb. Data are mean ± SEM (n = 4).

    Journal: PLoS ONE

    Article Title: ECIS technology reveals that monocytes isolated by CD14+ve selection mediate greater loss of BBB integrity than untouched monocytes, which occurs to a greater extent with IL-1β activated endothelium in comparison to TNFα

    doi: 10.1371/journal.pone.0180267

    Figure Lengend Snippet: Analysis of barrier formation in endothelial monolayer. (A) Expression of endothelial junctional molecules CD31, CD144 and ZO-1 (from left to right) and actin (far right) in hCMVEC monolayer 48 h after seeding. (B) The permeability of hCMVEC monolayer to FITC-dextran 48 h after seeding (mean ± SEM, n = 4). (C) Time course analysis of endothelial tight junction molecule occludin expression at 24, 48 and 72 h post seeding of hCMVECs using Western blot using a mouse monoclonal anti-occludin (left) and rabbit polyclonal anti-occludin (right). (D) Real time measurement of electrical resistance across the endothelial monolayer as an indicator of endothelial barrier integrity. The barrier resistance (Rb) and the basolateral adhesion (Alpha) due to focal adhesions can be modelled using the ECIS Z theta software when the experiment is run using multi-frequency mode (250 to 64000 Hz). The modelling reveals that the basolateral adhesion occurs very fast and junction formation begins around 10 hours after seeding. Overall barrier resistance is the summation of both Rb and alpha, but this software function can determine whether barrier changes are predominantly due to the Rb. Data are mean ± SEM (n = 4).

    Article Snippet: The blot was probed with a mouse monoclonal anti-occludin (cat# MAB7074, R & D systems, 1:400) or a rabbit polyclonal anti-occludin (cat# 71–1500, Invitrogen, 1:400) in immuno-buffer (2% bovine serum albumin in TBST) overnight at 4°C.

    Techniques: Expressing, Permeability, Western Blot, Electric Cell-substrate Impedance Sensing, Software

    Tight Junction Protein, Occludin, Expression in the Cerebral Vasculature of C57BL/6 Mice is not Altered through Exposure to Mixed Vehicle Emissions Double immunofluorescence of occludin (red) and vWF (green) in cerebral microvessels from 4 mo old C57BL/6 mice on a low-fat (LF; A–C) or high-fat diet (HF; G–I) exposed filtered air (FA) or on a LF (D–F) or HF diet (J–L) and to exposed MVE at 100 µg/m 3 PM of mixed gasoline and diesel engine emissions for 6hr/d, 7 d/wk, for 30 d (n=3–4 per exposure group; 4 sections each, 3–5 vessels per section). Right panels (yellow; C, F, I, L) are merged figures of left (red; A, D, G, J) and center (green, E, H, K) panels for occludin and vWF. Blue fluorescence is Hoechst stained nuclei. Scale bar = 100 µm. Results shown are mean ± SEM.

    Journal: Environmental research

    Article Title: Exposure to Traffic-Generated Air Pollutants Mediates Alterations in Brain Microvascular Integrity in Wildtype Mice on a High-Fat Diet

    doi: 10.1016/j.envres.2017.10.029

    Figure Lengend Snippet: Tight Junction Protein, Occludin, Expression in the Cerebral Vasculature of C57BL/6 Mice is not Altered through Exposure to Mixed Vehicle Emissions Double immunofluorescence of occludin (red) and vWF (green) in cerebral microvessels from 4 mo old C57BL/6 mice on a low-fat (LF; A–C) or high-fat diet (HF; G–I) exposed filtered air (FA) or on a LF (D–F) or HF diet (J–L) and to exposed MVE at 100 µg/m 3 PM of mixed gasoline and diesel engine emissions for 6hr/d, 7 d/wk, for 30 d (n=3–4 per exposure group; 4 sections each, 3–5 vessels per section). Right panels (yellow; C, F, I, L) are merged figures of left (red; A, D, G, J) and center (green, E, H, K) panels for occludin and vWF. Blue fluorescence is Hoechst stained nuclei. Scale bar = 100 µm. Results shown are mean ± SEM.

    Article Snippet: Frozen brain sections (10 µm) of the cerebrum (between Bregma 0 through −2.5mm) were prepared for either LOX-1, CD-36, 4-HNE, occludin, claudin-5, MMP-9, and/or double immunofluorescence, using techniques previously described by our laboratory ( ; ) with the following primary antibodies: MMP-9 (1:1000; Abcam, Cambridge, MA; Cat. #38898), occludin (1:500; Abcam #168986), claudin-5 (1:500; Abcam #15106), LOX-1 (1:1000; Abcam #60178), CD-36 (1:1000; Abcam #64014), 4-HNE (1:1000; Abcam #46545), AT-1 (1:250; Abcam #18801) and von Willebrand factor (vWF) (1:1000; Abcam #11713) and anti-rabbit Alexa Fluor 555 (1:500) and anti-sheep Alexa Fluor 455 (1:500) secondary antibodies.

    Techniques: Expressing, Mouse Assay, Immunofluorescence, Fluorescence, Staining

    The effects of L. frumenti on the intestinal epithelial physical barrier functions in early-weaned piglets. (A,B) Western blotting measurements of the expression levels of tight junction proteins (including ZO-1, Occludin, JAM-1, and Claudin-1) and a housekeeping protein (β-actin) in the small intestinal tissues (including duodenum, jejunum, and ileum) from early-weaned piglets (A) . Normalization and quantitation of ZO-1/β-actin, Occludin/β-actin, JAM-1/β-actin, and Claudin-1/β-actin as prepared described in (B) . Data are shown as mean ± SEM for at least three different experiments (ns, not significant, p > 0.05, ∗ p

    Journal: Frontiers in Microbiology

    Article Title: Lactobacillus frumenti Facilitates Intestinal Epithelial Barrier Function Maintenance in Early-Weaned Piglets

    doi: 10.3389/fmicb.2018.00897

    Figure Lengend Snippet: The effects of L. frumenti on the intestinal epithelial physical barrier functions in early-weaned piglets. (A,B) Western blotting measurements of the expression levels of tight junction proteins (including ZO-1, Occludin, JAM-1, and Claudin-1) and a housekeeping protein (β-actin) in the small intestinal tissues (including duodenum, jejunum, and ileum) from early-weaned piglets (A) . Normalization and quantitation of ZO-1/β-actin, Occludin/β-actin, JAM-1/β-actin, and Claudin-1/β-actin as prepared described in (B) . Data are shown as mean ± SEM for at least three different experiments (ns, not significant, p > 0.05, ∗ p

    Article Snippet: The following antibodies were used in the immunohistochemistry assay: Claudin-1 antibody (Abcam, ab211737), Occludin antibody (Abcam, ab216327), and ZO-1 antibody (Abcam, ab214228).

    Techniques: Western Blot, Expressing, Quantitation Assay

    Metformin attenuates Staphylococcus aureus induced cleavage of occludin at 60 kD and alters cleavage fragments at 44 kD. (A) Occludin bands were normalized to β‐actin and to their respective controls,* P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Metformin attenuates the effect of Staphylococcus aureus on airway tight junctions by increasing PKCζ‐mediated phosphorylation of occludin, et al. Metformin attenuates the effect of Staphylococcus aureus on airway tight junctions by increasing PKCζ‐mediated phosphorylation of occludin

    doi: 10.1111/jcmm.13929

    Figure Lengend Snippet: Metformin attenuates Staphylococcus aureus induced cleavage of occludin at 60 kD and alters cleavage fragments at 44 kD. (A) Occludin bands were normalized to β‐actin and to their respective controls,* P

    Article Snippet: Occludin antibody was a rabbit polyclonal used at (1:100 dilution) from Santa Cruz Biotechnology (H‐279, Wembley, UK).

    Techniques:

    Expression and cellular distribution of tight junction-associated protein, occludin, in septic plasma-stimulated MCVEC. Confluent MCVEC monolayers were stimulated with 20% (vol/vol) blood plasma obtained from sham ( lanes 1 , 2 , and 6 ) or FIP plasma

    Journal:

    Article Title: Role of endothelial nitric oxide synthase-derived nitric oxide in activation and dysfunction of cerebrovascular endothelial cells during early onsets of sepsis

    doi: 10.1152/ajpheart.00476.2008

    Figure Lengend Snippet: Expression and cellular distribution of tight junction-associated protein, occludin, in septic plasma-stimulated MCVEC. Confluent MCVEC monolayers were stimulated with 20% (vol/vol) blood plasma obtained from sham ( lanes 1 , 2 , and 6 ) or FIP plasma

    Article Snippet: The membranes were blocked overnight with 5% skim milk and then further incubated for 4 h at the room temperature with polyclonal antibody directed against occludin (clone H-279; Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Expressing

    FICZ prevents DSS-induced disruption of TJ. (A) Expressions of TJ proteins (ZO-1, Occludin and Claudin-1) and CYP1A1 in the colonic mucosa were evaluated by western blotting. (B) Immunohistochemical analysis of TJ proteins in the colonic mucosa. Magnification: ×400.

    Journal: International Journal of Biological Sciences

    Article Title: Aryl Hydrocarbon Receptor Activation Modulates Intestinal Epithelial Barrier Function by Maintaining Tight Junction Integrity

    doi: 10.7150/ijbs.22259

    Figure Lengend Snippet: FICZ prevents DSS-induced disruption of TJ. (A) Expressions of TJ proteins (ZO-1, Occludin and Claudin-1) and CYP1A1 in the colonic mucosa were evaluated by western blotting. (B) Immunohistochemical analysis of TJ proteins in the colonic mucosa. Magnification: ×400.

    Article Snippet: The sections were then blocked with 5% bovine serum albumin (BSA) at room temperature for 30 min and were subsequently incubated with the following primary antibodies overnight at 4 °C: ZO-1 (1:50, Proteintech), Occludin (1:50, Proteintech), Claudin-1 (1:50, Proteintech), MLCK (1:50, Proteintech) and pMLC (1:100, Abcam).

    Techniques: Western Blot, Immunohistochemistry

    Overall working model HHcy induced oxidative and ER stress conditions in the eyes of CBS +/− mice indicating the presence of ROS through the upregulation of BiP, PERK and downregulation of TJPs such as occludin respectively. These metabolic changes are generally accompanied by the excess production of glutamate. Such physiological alterations can contribute to the downregulation TJPs making blood vessels permeable which can lead to the loss of ocular homeostasis followed by vision impairment. GYY4137 treatment could potentially mitigate these harmful effects.

    Journal: International Journal of Ophthalmology

    Article Title: Hydrogen sulfide intervention in cystathionine-β-synthase mutant mouse helps restore ocular homeostasis

    doi: 10.18240/ijo.2019.05.09

    Figure Lengend Snippet: Overall working model HHcy induced oxidative and ER stress conditions in the eyes of CBS +/− mice indicating the presence of ROS through the upregulation of BiP, PERK and downregulation of TJPs such as occludin respectively. These metabolic changes are generally accompanied by the excess production of glutamate. Such physiological alterations can contribute to the downregulation TJPs making blood vessels permeable which can lead to the loss of ocular homeostasis followed by vision impairment. GYY4137 treatment could potentially mitigate these harmful effects.

    Article Snippet: Antibodies for CBS, CSE, and glyceraldehyde 3 phosphate dehydrogenase (GAPDH) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), MTHFR, Hcy and superoxide dismutase 1 (SOD1) from Abcam (Cambridge, MA, USA), BiP, protein kinase R like ER kinase (PERK), CAAT enhancer binding protein homologous protein (CHOP), AMP activated protein kinase (AMPK), p-AMPK from Cell Signaling Technology (Danvers, MA, USA), GAPDH was from Boster Biological Technology (Pleasanton, CA, USA) and occludin was purchased from Novus Biologicals (Littleton, CO, USA).

    Techniques: Mouse Assay

    Angiographic imaging of mice eyes and their retinae A: HHcy induced microvasculature changes in mice eyes (imaging with barium sulphate). B: Quantitation of vessels' density from A. C: BSA-FITC images showing leakage of vessels (arrow) as observed on retinal flat mounts of CBS +/− treated mice group. GYY4137 could halt leakage of the retinal vasculature in CBS +/− mice. D: Graph represents the percent change in vascular density in different mice groups. E: Decreased expression of TJP occludin in the CBS +/− mice was rescued by GYY4137 treatment. F: The bar graph showing the quantitative estimation of occludin after normalization with GAPDH, data presented as mean±SEM. a P

    Journal: International Journal of Ophthalmology

    Article Title: Hydrogen sulfide intervention in cystathionine-β-synthase mutant mouse helps restore ocular homeostasis

    doi: 10.18240/ijo.2019.05.09

    Figure Lengend Snippet: Angiographic imaging of mice eyes and their retinae A: HHcy induced microvasculature changes in mice eyes (imaging with barium sulphate). B: Quantitation of vessels' density from A. C: BSA-FITC images showing leakage of vessels (arrow) as observed on retinal flat mounts of CBS +/− treated mice group. GYY4137 could halt leakage of the retinal vasculature in CBS +/− mice. D: Graph represents the percent change in vascular density in different mice groups. E: Decreased expression of TJP occludin in the CBS +/− mice was rescued by GYY4137 treatment. F: The bar graph showing the quantitative estimation of occludin after normalization with GAPDH, data presented as mean±SEM. a P

    Article Snippet: Antibodies for CBS, CSE, and glyceraldehyde 3 phosphate dehydrogenase (GAPDH) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), MTHFR, Hcy and superoxide dismutase 1 (SOD1) from Abcam (Cambridge, MA, USA), BiP, protein kinase R like ER kinase (PERK), CAAT enhancer binding protein homologous protein (CHOP), AMP activated protein kinase (AMPK), p-AMPK from Cell Signaling Technology (Danvers, MA, USA), GAPDH was from Boster Biological Technology (Pleasanton, CA, USA) and occludin was purchased from Novus Biologicals (Littleton, CO, USA).

    Techniques: Imaging, Mouse Assay, Quantitation Assay, Expressing

    Effects of resveratrol on H 2 O 2 -induced cytotoxicity in Nrf2-knockdown IPEC-J2 cells. The Nrf2-knockdown (shNRF2-IPEC-J2) or control (SC-IPEC-J2) cells were pretreated with or without 50 μ M RSV for 6 hours and then cocultured with 500 μ M H 2 O 2 for 4 hours. (a) Cell viability was measured using the CCK-8 assay. Results are presented as the percentage of cell viability compared with the control (0 μ M). (b, c) The relative expression of SOD-1, CAT, claudin-1, occludin, and ZO-1 was detected by qRT-PCR. Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (d) Apoptotic cells were analyzed by flow cytometry using Annexin V-PI double staining. (e) Protein levels of Nrf2, Akt, p-Nrf2, and p-Akt were detected by Western blot with β -actin as the loading control. Values are the mean ± SE. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Resveratrol Attenuates Oxidative Stress-Induced Intestinal Barrier Injury through PI3K/Akt-Mediated Nrf2 Signaling Pathway

    doi: 10.1155/2019/7591840

    Figure Lengend Snippet: Effects of resveratrol on H 2 O 2 -induced cytotoxicity in Nrf2-knockdown IPEC-J2 cells. The Nrf2-knockdown (shNRF2-IPEC-J2) or control (SC-IPEC-J2) cells were pretreated with or without 50 μ M RSV for 6 hours and then cocultured with 500 μ M H 2 O 2 for 4 hours. (a) Cell viability was measured using the CCK-8 assay. Results are presented as the percentage of cell viability compared with the control (0 μ M). (b, c) The relative expression of SOD-1, CAT, claudin-1, occludin, and ZO-1 was detected by qRT-PCR. Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (d) Apoptotic cells were analyzed by flow cytometry using Annexin V-PI double staining. (e) Protein levels of Nrf2, Akt, p-Nrf2, and p-Akt were detected by Western blot with β -actin as the loading control. Values are the mean ± SE. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Article Snippet: The antibodies against Nrf2, Keap1, and β -actin were obtained from Abcam, the antibodies against claudin-1 and occludin from Selleckchem (Houston, United States), and the antibody against ZO-1 from Proteintech (Wuhan, China).

    Techniques: CCK-8 Assay, Expressing, Quantitative RT-PCR, Flow Cytometry, Cytometry, Double Staining, Western Blot

    Effects of RSV and the selective inhibitor LY294002 on H 2 O 2 -induced cytotoxicity in IPEC-J2 cells. IPEC-J2 cells were pretreated with 25 μ M LY294002 for 2 h and then treated with 50 μ M RSV for 4 h before incubation with 500 μ M H 2 O 2 for 4 hours. (a) Cell viability was measured using the CCK-8 assay. Results are presented as the percentage of cell viability compared with the control (0 μ M). (b, c) The relative expression of SOD-1, CAT, claudin-1, occludin, and ZO-1 was detected by qRT-PCR. Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (d) The apoptotic cells were analyzed by flow cytometry using Annexin V-PI double staining. (e) Protein levels of Nrf2, Akt, p-Nrf2, and p-Akt were detected by Western blot with β -actin as the loading control. Values are the mean ± SE. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Resveratrol Attenuates Oxidative Stress-Induced Intestinal Barrier Injury through PI3K/Akt-Mediated Nrf2 Signaling Pathway

    doi: 10.1155/2019/7591840

    Figure Lengend Snippet: Effects of RSV and the selective inhibitor LY294002 on H 2 O 2 -induced cytotoxicity in IPEC-J2 cells. IPEC-J2 cells were pretreated with 25 μ M LY294002 for 2 h and then treated with 50 μ M RSV for 4 h before incubation with 500 μ M H 2 O 2 for 4 hours. (a) Cell viability was measured using the CCK-8 assay. Results are presented as the percentage of cell viability compared with the control (0 μ M). (b, c) The relative expression of SOD-1, CAT, claudin-1, occludin, and ZO-1 was detected by qRT-PCR. Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (d) The apoptotic cells were analyzed by flow cytometry using Annexin V-PI double staining. (e) Protein levels of Nrf2, Akt, p-Nrf2, and p-Akt were detected by Western blot with β -actin as the loading control. Values are the mean ± SE. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Article Snippet: The antibodies against Nrf2, Keap1, and β -actin were obtained from Abcam, the antibodies against claudin-1 and occludin from Selleckchem (Houston, United States), and the antibody against ZO-1 from Proteintech (Wuhan, China).

    Techniques: Incubation, CCK-8 Assay, Expressing, Quantitative RT-PCR, Flow Cytometry, Cytometry, Double Staining, Western Blot

    Effects of resveratrol and/or H 2 O 2 on the levels of TJ proteins. IPEC-J2 cells were pretreated with the indicated concentrations of RSV and then cocultured with 500 μ M H 2 O 2 for 4 hours. (a) The expression of claudin-1, occludin, and ZO-1 was detected by quantitative real-time PCR (qRT-PCR). Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (b) Protein levels of claudin-1, occludin, and ZO-1 were detected by Western blot with β -actin as the loading control. Values are the mean ± SE; n = 3. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Resveratrol Attenuates Oxidative Stress-Induced Intestinal Barrier Injury through PI3K/Akt-Mediated Nrf2 Signaling Pathway

    doi: 10.1155/2019/7591840

    Figure Lengend Snippet: Effects of resveratrol and/or H 2 O 2 on the levels of TJ proteins. IPEC-J2 cells were pretreated with the indicated concentrations of RSV and then cocultured with 500 μ M H 2 O 2 for 4 hours. (a) The expression of claudin-1, occludin, and ZO-1 was detected by quantitative real-time PCR (qRT-PCR). Data are shown as ratios of abundance of target gene transcripts in the treated cells to those in the control cells after normalization to β -actin. (b) Protein levels of claudin-1, occludin, and ZO-1 were detected by Western blot with β -actin as the loading control. Values are the mean ± SE; n = 3. The columns with the same superscript capital letters and with different superscript capital letters mean no significant difference ( P > 0.05) and significant difference ( P

    Article Snippet: The antibodies against Nrf2, Keap1, and β -actin were obtained from Abcam, the antibodies against claudin-1 and occludin from Selleckchem (Houston, United States), and the antibody against ZO-1 from Proteintech (Wuhan, China).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

    Immunohistochemical expression of occludin in the colonic mucosa of patients with (a) non-CKD, (b) advanced CKD, and (c) HD with hypotension. The total of epithelial cells lining villi exhibited positive immunostaining for occludin in healthy controls (a). In patients with advanced CKD, occludin expression is reduced in numerous epithelial cells (b). In HD patients with IDH, there was profound loss of occludin expression by most epithelial cells lining villi (blue arrows) (c).

    Journal: BioMed Research International

    Article Title: Impaired Gut Epithelial Tight Junction Expression in Hemodialysis Patients Complicated with Intradialytic Hypotension

    doi: 10.1155/2018/2670312

    Figure Lengend Snippet: Immunohistochemical expression of occludin in the colonic mucosa of patients with (a) non-CKD, (b) advanced CKD, and (c) HD with hypotension. The total of epithelial cells lining villi exhibited positive immunostaining for occludin in healthy controls (a). In patients with advanced CKD, occludin expression is reduced in numerous epithelial cells (b). In HD patients with IDH, there was profound loss of occludin expression by most epithelial cells lining villi (blue arrows) (c).

    Article Snippet: Claudin-1 antibody, claudin-4 antibody, occludin antibody, and Zona occludens-1 (ZO-1) antibody were purchased from Biorbyt Company (Cambridge, Cambridgeshire, United Kingdom).

    Techniques: Immunohistochemistry, Expressing, Immunostaining

    Raft localization of occludin. Caco-2 cells were untreated (A), treated with CPE (2.5 μg/ml) for 60 min (B), or treated for 60 min with 10 mM MβCD (C), followed by cold Triton X-100 extraction and sucrose density gradient centrifugation.

    Journal:

    Article Title: Evidence that Membrane Rafts Are Not Required for the Action of Clostridium perfringens Enterotoxin ▿

    doi: 10.1128/IAI.00854-08

    Figure Lengend Snippet: Raft localization of occludin. Caco-2 cells were untreated (A), treated with CPE (2.5 μg/ml) for 60 min (B), or treated for 60 min with 10 mM MβCD (C), followed by cold Triton X-100 extraction and sucrose density gradient centrifugation.

    Article Snippet: Claudin-4 and occludin antibodies were purchased from Invitrogen (Carlsbad, CA).

    Techniques: Gradient Centrifugation

    Effects of SC and CH-1 on occludin distribution. (A) After treatment with CPE (2.5 μg/ml) for 15 min at 4°C, Caco-2 cells were extracted with cold Triton X-100 and subjected to sucrose gradient centrifugation. Gradient fractions 2 to 5

    Journal:

    Article Title: Evidence that Membrane Rafts Are Not Required for the Action of Clostridium perfringens Enterotoxin ▿

    doi: 10.1128/IAI.00854-08

    Figure Lengend Snippet: Effects of SC and CH-1 on occludin distribution. (A) After treatment with CPE (2.5 μg/ml) for 15 min at 4°C, Caco-2 cells were extracted with cold Triton X-100 and subjected to sucrose gradient centrifugation. Gradient fractions 2 to 5

    Article Snippet: Claudin-4 and occludin antibodies were purchased from Invitrogen (Carlsbad, CA).

    Techniques: Gradient Centrifugation

    Kinetics of CPE complex formation and occludin distribution. (A) Caco-2 monolayers were treated with 2.5 μg/ml of CPE for the indicated times (10, 30, and 45 min), and sucrose gradients were then performed for Triton X-100 lysates of each time.

    Journal:

    Article Title: Evidence that Membrane Rafts Are Not Required for the Action of Clostridium perfringens Enterotoxin ▿

    doi: 10.1128/IAI.00854-08

    Figure Lengend Snippet: Kinetics of CPE complex formation and occludin distribution. (A) Caco-2 monolayers were treated with 2.5 μg/ml of CPE for the indicated times (10, 30, and 45 min), and sucrose gradients were then performed for Triton X-100 lysates of each time.

    Article Snippet: Claudin-4 and occludin antibodies were purchased from Invitrogen (Carlsbad, CA).

    Techniques:

    Nectin-2, but not occludin, is expressed in RV, comma-shaped bodies, and S-shaped bodies. Five-micrometer sections of e18 mouse kidney were stained with monoclonal anti-nectin-2 antibody ( A ′, A ′′′, and B – E ) and polyclonal anti-occludin antibody ( A , A ′′′, and B – E ), Hoechst 33342 ( A ′′, A ′′′, and B – D ), and lotus lectin (LTL; blue staining, E ). UB and RV are labeled using arrows. Open arrowheads demonstrate punctate nectin-2 and occludin staining in UB ( A – A ′′′). In B at high magnification, yellow indicates colocalization of nectin-2 and occludin, and green and red indicate areas where nectin-2 and occludin did not colocalize (open arrowheads and filled arrowheads, respectively). Asterisks demonstrate RV, comma-shaped body, and S-shaped body where nectin-2 is expressed, but occludin is not ( A – A ′′′, C , and D ). Scale bars = 10 μm.

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Nectin proteins are expressed at early stages of nephrogenesis and play a role in renal epithelial cell morphogenesis

    doi: 10.1152/ajprenal.90328.2008

    Figure Lengend Snippet: Nectin-2, but not occludin, is expressed in RV, comma-shaped bodies, and S-shaped bodies. Five-micrometer sections of e18 mouse kidney were stained with monoclonal anti-nectin-2 antibody ( A ′, A ′′′, and B – E ) and polyclonal anti-occludin antibody ( A , A ′′′, and B – E ), Hoechst 33342 ( A ′′, A ′′′, and B – D ), and lotus lectin (LTL; blue staining, E ). UB and RV are labeled using arrows. Open arrowheads demonstrate punctate nectin-2 and occludin staining in UB ( A – A ′′′). In B at high magnification, yellow indicates colocalization of nectin-2 and occludin, and green and red indicate areas where nectin-2 and occludin did not colocalize (open arrowheads and filled arrowheads, respectively). Asterisks demonstrate RV, comma-shaped body, and S-shaped body where nectin-2 is expressed, but occludin is not ( A – A ′′′, C , and D ). Scale bars = 10 μm.

    Article Snippet: PAR-3 (Millipore, Billerica, MA), β-catenin (Santa Cruz Biotechnology, Santa Cruz, CA), and occludin (BD Biosciences) antibodies were purchased.

    Techniques: Staining, Labeling

    KO of LRRC75A-AS1 enhanced TJ of MAC-T cells. (A) Gene expressions of Claudin-1, Occludin and ZO-1 were analyzed by RT-qPCR. (B) Claudin-1, Occludin and ZO-1 expression in KO-Control and KO-LRRC75A-AS1 cells were analyzed by western blot. GAPDH was used as an internal control. (C) The relative quantitative analysis of western blot result was performed. *P

    Journal: International Journal of Biological Sciences

    Article Title: LRRC75A antisense lncRNA1 knockout attenuates inflammatory responses of bovine mammary epithelial cells

    doi: 10.7150/ijbs.38214

    Figure Lengend Snippet: KO of LRRC75A-AS1 enhanced TJ of MAC-T cells. (A) Gene expressions of Claudin-1, Occludin and ZO-1 were analyzed by RT-qPCR. (B) Claudin-1, Occludin and ZO-1 expression in KO-Control and KO-LRRC75A-AS1 cells were analyzed by western blot. GAPDH was used as an internal control. (C) The relative quantitative analysis of western blot result was performed. *P

    Article Snippet: After resolved proteins were blotted onto PVDF transfer membranes and blocked with 10% non-fat milk in TBST for 2 h, the membranes were incubated with primary antibodies against ZO-1 (Bioss, Beijing, China), Occludin (Bioss), Claudin-1 (Bioss), p65 (Santa Cruz, Dallas, TX, USA), phosphorylated p65 (Bioss), GAPDH (Bioss) at 4°C overnight.

    Techniques: Quantitative RT-PCR, Expressing, Western Blot

    Lower expression of LRRC75A enhanced TJ and weakened the activation of inflammation. (A) LRRC75A was detected at indicated time point by RT-qPCR after siRNA was transfected into cells. (B) LRRC75A was overexpressed by CD513B-1 plasmid and detected by RT-qPCR. (C) and (D) The mRNA levels of Claudin-1, Occludin and ZO-1 were detected by RT-qPCR after siRNA and overexpression plasmid were respectively transfected into cells. (E) TJ proteins were detected by western blot after siRNA and overexpression plasmid were respectively transfected into cell. (F) Relative quantative analyze of TJ proteins in western blot was performed. (G) P65 in the nucleus and total P-p65 was detected by western blot. H3 was used as control in nucleus. GAPDH was regarded as control in cytoplasm. (H) Relative quantative analyze of p65 in the nucleus and P-p65 in western blot was performed. (I) Activation of NF-κB pathway was measured by dual luciferase reporter gene assay. (NC: Negative Control; Si: SiRNA knockdown; CD513B-1: Empty vector; Over: Overexpression.)

    Journal: International Journal of Biological Sciences

    Article Title: LRRC75A antisense lncRNA1 knockout attenuates inflammatory responses of bovine mammary epithelial cells

    doi: 10.7150/ijbs.38214

    Figure Lengend Snippet: Lower expression of LRRC75A enhanced TJ and weakened the activation of inflammation. (A) LRRC75A was detected at indicated time point by RT-qPCR after siRNA was transfected into cells. (B) LRRC75A was overexpressed by CD513B-1 plasmid and detected by RT-qPCR. (C) and (D) The mRNA levels of Claudin-1, Occludin and ZO-1 were detected by RT-qPCR after siRNA and overexpression plasmid were respectively transfected into cells. (E) TJ proteins were detected by western blot after siRNA and overexpression plasmid were respectively transfected into cell. (F) Relative quantative analyze of TJ proteins in western blot was performed. (G) P65 in the nucleus and total P-p65 was detected by western blot. H3 was used as control in nucleus. GAPDH was regarded as control in cytoplasm. (H) Relative quantative analyze of p65 in the nucleus and P-p65 in western blot was performed. (I) Activation of NF-κB pathway was measured by dual luciferase reporter gene assay. (NC: Negative Control; Si: SiRNA knockdown; CD513B-1: Empty vector; Over: Overexpression.)

    Article Snippet: After resolved proteins were blotted onto PVDF transfer membranes and blocked with 10% non-fat milk in TBST for 2 h, the membranes were incubated with primary antibodies against ZO-1 (Bioss, Beijing, China), Occludin (Bioss), Claudin-1 (Bioss), p65 (Santa Cruz, Dallas, TX, USA), phosphorylated p65 (Bioss), GAPDH (Bioss) at 4°C overnight.

    Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Transfection, Plasmid Preparation, Over Expression, Western Blot, Luciferase, Reporter Gene Assay, Negative Control