Structured Review

Abcam occludin
Confirmation of ZO-1, <t>occludin</t> and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin
Occludin, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/occludin/product/Abcam
Average 99 stars, based on 153 article reviews
Price from $9.99 to $1999.99
occludin - by Bioz Stars, 2020-08
99/100 stars

Images

1) Product Images from "Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction"

Article Title: Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

Journal: Molecular Medicine

doi: 10.2119/molmed.2011.00107

Confirmation of ZO-1, occludin and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin
Figure Legend Snippet: Confirmation of ZO-1, occludin and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin

Techniques Used: Quantitative RT-PCR

Decreased Expression of ZO-1, Occludin and Claudin-1 in STR-IO As Confirmed by QRT-PCR, Western Blotting and IHC
Figure Legend Snippet: Decreased Expression of ZO-1, Occludin and Claudin-1 in STR-IO As Confirmed by QRT-PCR, Western Blotting and IHC

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

2) Product Images from "Exposure to Traffic-Generated Air Pollutants Mediates Alterations in Brain Microvascular Integrity in Wildtype Mice on a High-Fat Diet"

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

Journal: Environmental research

doi: 10.1016/j.envres.2017.10.029

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.
Figure Legend 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.

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

3) Product Images from "Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis"

Article Title: Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis

Journal: Nature Communications

doi: 10.1038/s41467-018-07810-w

CRC-secreted miR-25-3p primes pre-metastatic niche. a Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular permeability of mice liver by in vivo permeability assay. The mice were injected with rhodamine–dextran after exposure to PKH67-labeled exosomes. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided ( n = 5). Scale bar represents 50 µm. b Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular KLF4, KLF2, and ZO-1 expression (red) in hepatic vessels by immunofluorescence. The vascular structures were labeled by CD34 (green). Scale bar represents 50 µm. c Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on KLF4, ZO-1, occludin, Claudin5, KLF2, VEGFR2, fibronectin, and S100 expression in mice liver by Western blot. d The mice were intra-spleen injected with naked SW480 cells after exposure to NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments. The number of liver metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided ( n = 5). Scale bar in left panels represents 0.5 cm. Scale bar in right panels represents 100 µm. ** P
Figure Legend Snippet: CRC-secreted miR-25-3p primes pre-metastatic niche. a Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular permeability of mice liver by in vivo permeability assay. The mice were injected with rhodamine–dextran after exposure to PKH67-labeled exosomes. Levels of rhodamine–dextran fluorescence in tissues were quantified using Image J software and normalized to the levels of DAPI. Mean ± SEM are provided ( n = 5). Scale bar represents 50 µm. b Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on vascular KLF4, KLF2, and ZO-1 expression (red) in hepatic vessels by immunofluorescence. The vascular structures were labeled by CD34 (green). Scale bar represents 50 µm. c Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments on KLF4, ZO-1, occludin, Claudin5, KLF2, VEGFR2, fibronectin, and S100 expression in mice liver by Western blot. d The mice were intra-spleen injected with naked SW480 cells after exposure to NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes or SW480/miR-25-3p exosomes + miR-25-3p inhibitor treatments. The number of liver metastatic sites (indicated by arrows) was counted under the microscope. Mean ± SEM are provided ( n = 5). Scale bar in left panels represents 0.5 cm. Scale bar in right panels represents 100 µm. ** P

Techniques Used: Permeability, Mouse Assay, In Vivo, Injection, Labeling, Fluorescence, Software, Expressing, Immunofluorescence, Western Blot, Microscopy

CRC-secreted miR-25-3p silences KLF2 and KLF4 in HUVECs. a Western blot analysis of KLF2, VEGFR2, AKT, p-AKT, ERK, p-ERK expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 groups. b Western blot analysis of KLF4, ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. c Immunofluorescence staining analysis of ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. Scale bars represents 10 µm. d Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). e Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 treatments on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). f Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided ( n = 3). ** P
Figure Legend Snippet: CRC-secreted miR-25-3p silences KLF2 and KLF4 in HUVECs. a Western blot analysis of KLF2, VEGFR2, AKT, p-AKT, ERK, p-ERK expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 groups. b Western blot analysis of KLF4, ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. c Immunofluorescence staining analysis of ZO-1, occludin, Claudin5 expression in HUVECs incubated with NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 groups. Scale bars represents 10 µm. d Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). e Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF2 treatments on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). f Effects of NCM460 exosomes, SW480/mock exosomes, SW480/miR-25-3p exosomes, SW480/miR-25-3p exosomes + Annexin V, SW480/miR-25-3p exosomes + miR-25-3p inhibitor and SW480/miR-25-3p exosomes + KLF4 treatments on permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided ( n = 3). ** P

Techniques Used: Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Tube Formation Assay, Permeability, In Vitro

KLF2 and KLF4 are functional targets of miR-25-3p in HUVECs. a , b Luciferase activities of 3’UTR KLF2-luc and 3’UTR KLF4-luc constructs in HEK293A, HUVECs after transfection of miR-25-3p mimics. Mean ± SEM are provided ( n = 3). c KLF4, ZO-1, occludin, Claudin5 expression in miR-25-3p overexpressing or miR-25-3p/KLF4 co-expressing HUVECs by western blot. d KLF2, VEGFR2, AKT, p-AKT, ERK, p-ERK expression in miR-25-3p overexpressing or miR-25-3p/KLF2 co-expressing HUVECs by western blot. e Effects of miR-25-3p and miR-25-3p/KLF4 on permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided ( n = 3). f Effects of miR-25-3p and miR-25-3p/KLF2 on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). g Effects of miR-25-3p and miR-25-3p/KLF2 on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). h Effect of miR-25-3p knockdown on permeability of HUVEC monolayers by in vitro permeability assay. i Effect of miR-25-3p knockdown on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). j Effect of miR-25-3p knockdown on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). ** P
Figure Legend Snippet: KLF2 and KLF4 are functional targets of miR-25-3p in HUVECs. a , b Luciferase activities of 3’UTR KLF2-luc and 3’UTR KLF4-luc constructs in HEK293A, HUVECs after transfection of miR-25-3p mimics. Mean ± SEM are provided ( n = 3). c KLF4, ZO-1, occludin, Claudin5 expression in miR-25-3p overexpressing or miR-25-3p/KLF4 co-expressing HUVECs by western blot. d KLF2, VEGFR2, AKT, p-AKT, ERK, p-ERK expression in miR-25-3p overexpressing or miR-25-3p/KLF2 co-expressing HUVECs by western blot. e Effects of miR-25-3p and miR-25-3p/KLF4 on permeability of HUVEC monolayers by in vitro permeability assay. Mean ± SEM are provided ( n = 3). f Effects of miR-25-3p and miR-25-3p/KLF2 on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). g Effects of miR-25-3p and miR-25-3p/KLF2 on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). h Effect of miR-25-3p knockdown on permeability of HUVEC monolayers by in vitro permeability assay. i Effect of miR-25-3p knockdown on vascular outgrowth of rat aortic rings. Scale bar represents 200 µm. Mean ± SEM are provided ( n = 3). j Effect of miR-25-3p knockdown on tube formation ability of HUVECs by tube formation assay. Scale bar represents 100 µm. Mean ± SEM are provided ( n = 3). ** P

Techniques Used: Functional Assay, Luciferase, Construct, Transfection, Expressing, Western Blot, Permeability, In Vitro, Tube Formation Assay

4) Product Images from "Blood-brain barrier disruption induced by diagnostic ultrasound combined with microbubbles in mice"

Article Title: Blood-brain barrier disruption induced by diagnostic ultrasound combined with microbubbles in mice

Journal: Oncotarget

doi: 10.18632/oncotarget.23527

Representative blots ( A ) and relative quantitative analysis ( B , C , D ) of TJ related proteins ZO-1, occludin and claudin-5 expression in each group. Data were shown as the mean ± SEM, * P
Figure Legend Snippet: Representative blots ( A ) and relative quantitative analysis ( B , C , D ) of TJ related proteins ZO-1, occludin and claudin-5 expression in each group. Data were shown as the mean ± SEM, * P

Techniques Used: Expressing

Distribution and expression level of TJ related proteins ZO-1 ( A ), occludin ( C ) and claudin-5 ( E ) observed via immunohistofluorescence staining in each group. Relative fluorescence intensity of ZO-1 ( B ), occludin ( D ) and claudin-5 ( F ) compare with the control group. Data were shown as the mean ± SEM, * P
Figure Legend Snippet: Distribution and expression level of TJ related proteins ZO-1 ( A ), occludin ( C ) and claudin-5 ( E ) observed via immunohistofluorescence staining in each group. Relative fluorescence intensity of ZO-1 ( B ), occludin ( D ) and claudin-5 ( F ) compare with the control group. Data were shown as the mean ± SEM, * P

Techniques Used: Expressing, Immunohistofluorescence, Staining, Fluorescence

5) Product Images from "Musca domestica Cecropin (Mdc) Alleviates Salmonella typhimurium-Induced Colonic Mucosal Barrier Impairment: Associating With Inflammatory and Oxidative Stress Response, Tight Junction as Well as Intestinal Flora"

Article Title: Musca domestica Cecropin (Mdc) Alleviates Salmonella typhimurium-Induced Colonic Mucosal Barrier Impairment: Associating With Inflammatory and Oxidative Stress Response, Tight Junction as Well as Intestinal Flora

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2019.00522

Real-time quantitative PCR detection of ZO-1 mRNA (A) , Claudin-1 mRNA (B) , Occludin mRNA (C) , P65 mRNA (D) , COX-2 mRNA (E) , and INOS mRNA (F) in colon of 4 groups. Values are means ± SEMs, n = 6. ∗∗ p ≤ 0.01, ∗∗∗∗ p ≤ 0.0001, ns, no significant difference.
Figure Legend Snippet: Real-time quantitative PCR detection of ZO-1 mRNA (A) , Claudin-1 mRNA (B) , Occludin mRNA (C) , P65 mRNA (D) , COX-2 mRNA (E) , and INOS mRNA (F) in colon of 4 groups. Values are means ± SEMs, n = 6. ∗∗ p ≤ 0.01, ∗∗∗∗ p ≤ 0.0001, ns, no significant difference.

Techniques Used: Real-time Polymerase Chain Reaction

Immunofluorescence analysis of Occludin and P65 in colon of 4 groups. Values are means ± SEMs, n = 3. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001.
Figure Legend Snippet: Immunofluorescence analysis of Occludin and P65 in colon of 4 groups. Values are means ± SEMs, n = 3. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001.

Techniques Used: Immunofluorescence

Western blot analysis of ZO-1,Claudin-1, Occludin, P65, COX-2, INOS protein expression in relative to GAPDH in the colon of 4 groups (A,B) . Values are means ± SEMs, n = 3. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001, ns = no significant difference.
Figure Legend Snippet: Western blot analysis of ZO-1,Claudin-1, Occludin, P65, COX-2, INOS protein expression in relative to GAPDH in the colon of 4 groups (A,B) . Values are means ± SEMs, n = 3. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001, ns = no significant difference.

Techniques Used: Western Blot, Expressing

6) Product Images from "Regulatory T Cells Restrict Permeability to Bacterial Antigen Translocation and Preserve Short‐Chain Fatty Acids in Experimental Cirrhosis"

Article Title: Regulatory T Cells Restrict Permeability to Bacterial Antigen Translocation and Preserve Short‐Chain Fatty Acids in Experimental Cirrhosis

Journal: Hepatology Communications

doi: 10.1002/hep4.1268

TJ protein expression in WT and Rag1 ‐/‐ cirrhotic mice study groups. (A) mRNA transcripts relative expression of ZO‐1, occludin, claudin‐1, and claudin‐2 in colon tissue homogenates. (B) Representative images of colonic tissue sections from the different study groups. Protein expression was blindly measured in user‐specified regions of interest (ROIs) as percent brown area in hematoxylin‐stained colon samples using the ImageJ software ( http: //rsbweb.nih.gov ). The mean and SDs from three independent ROIs in each sample are represented in bar graphs. Negative control hematoxylin‐stained sections without primary antibody are also shown. (C) Western blot analysis of ZO‐1, occludin, claudin‐1, and claudin‐2 in colon tissue homogenates. Band densitometry analysis shows the mean and SDs from four independent blots run for each protein. * P
Figure Legend Snippet: TJ protein expression in WT and Rag1 ‐/‐ cirrhotic mice study groups. (A) mRNA transcripts relative expression of ZO‐1, occludin, claudin‐1, and claudin‐2 in colon tissue homogenates. (B) Representative images of colonic tissue sections from the different study groups. Protein expression was blindly measured in user‐specified regions of interest (ROIs) as percent brown area in hematoxylin‐stained colon samples using the ImageJ software ( http: //rsbweb.nih.gov ). The mean and SDs from three independent ROIs in each sample are represented in bar graphs. Negative control hematoxylin‐stained sections without primary antibody are also shown. (C) Western blot analysis of ZO‐1, occludin, claudin‐1, and claudin‐2 in colon tissue homogenates. Band densitometry analysis shows the mean and SDs from four independent blots run for each protein. * P

Techniques Used: Expressing, Mouse Assay, Staining, Software, Negative Control, Western Blot

7) Product Images from "Omega-3 polyunsaturated fatty acids mitigate blood-brain barrier disruption after hypoxic-ischemic brain injury"

Article Title: Omega-3 polyunsaturated fatty acids mitigate blood-brain barrier disruption after hypoxic-ischemic brain injury

Journal: Neurobiology of disease

doi: 10.1016/j.nbd.2016.02.020

n-3 PUFAs preserve the integrity of tight junctions after neonatal H/I. ( A ) Western blots showing expression of occludin, ZO-1, claudin-5, and cadherin-10 at 48 hours after H/I. ( B ) Quantification of occludin, ZO-1, claudin-5, and cadherin-10 levels at 48 hours after H/I. **p≤0.01,*p≤0.05 vs Sham, ##p≤0.01, #p≤0.05 vs N3L H/I, data are mean±s.d., n=6 each group. ( C ) Representative electron microscopic images of neuronal ultrastructure and the components of the BBB at 48 hours after H/I. N: neuron; P: pericyte, EC: endothelial cells; BM: base membranes; T: tight junction; Lumen: capillary lumen; R: a red cell; A: perivascular astrocytes; M: mitochondria. Scale bar: from top to bottom rows: 5µm, 2µm, and 1µm. ( D ) Representative immunofluorescent images of occludin, ZO-1, and claudin-5 with RECA + vessels and DAPI nuclear labeling at 48 hours following H/I brain injury (scale bar=20µm).
Figure Legend Snippet: n-3 PUFAs preserve the integrity of tight junctions after neonatal H/I. ( A ) Western blots showing expression of occludin, ZO-1, claudin-5, and cadherin-10 at 48 hours after H/I. ( B ) Quantification of occludin, ZO-1, claudin-5, and cadherin-10 levels at 48 hours after H/I. **p≤0.01,*p≤0.05 vs Sham, ##p≤0.01, #p≤0.05 vs N3L H/I, data are mean±s.d., n=6 each group. ( C ) Representative electron microscopic images of neuronal ultrastructure and the components of the BBB at 48 hours after H/I. N: neuron; P: pericyte, EC: endothelial cells; BM: base membranes; T: tight junction; Lumen: capillary lumen; R: a red cell; A: perivascular astrocytes; M: mitochondria. Scale bar: from top to bottom rows: 5µm, 2µm, and 1µm. ( D ) Representative immunofluorescent images of occludin, ZO-1, and claudin-5 with RECA + vessels and DAPI nuclear labeling at 48 hours following H/I brain injury (scale bar=20µm).

Techniques Used: Western Blot, Expressing, Labeling

8) Product Images from "MnTE-2-PyP Attenuates TGF-β-Induced Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Inhibiting the Smad2/3 Signaling Pathway"

Article Title: MnTE-2-PyP Attenuates TGF-β-Induced Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Inhibiting the Smad2/3 Signaling Pathway

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2019/8639791

MnTE-2-PyP attenuates TGF- β -induced expression of EMT markers in colorectal cancer cells. (a) The SW480 cells were pretreated with MnTE-2-PyP (30 μ M) for 12 h and then treated with TGF- β (5 ng/ml) for 24 h. Western blotting analysis showed that TGF- β decreased the expression levels of the epithelial cell markers E-cadherin and occludin and increased the mesenchymal markers N-cadherin and vimentin in SW480 cells. MnTE-2-PyP suppressed the changes of markers related to EMT caused by TGF- β in SW480 cells. (b–f) Quantification of protein expression shown in (a) is normalized to GADPH. ∗ P
Figure Legend Snippet: MnTE-2-PyP attenuates TGF- β -induced expression of EMT markers in colorectal cancer cells. (a) The SW480 cells were pretreated with MnTE-2-PyP (30 μ M) for 12 h and then treated with TGF- β (5 ng/ml) for 24 h. Western blotting analysis showed that TGF- β decreased the expression levels of the epithelial cell markers E-cadherin and occludin and increased the mesenchymal markers N-cadherin and vimentin in SW480 cells. MnTE-2-PyP suppressed the changes of markers related to EMT caused by TGF- β in SW480 cells. (b–f) Quantification of protein expression shown in (a) is normalized to GADPH. ∗ P

Techniques Used: Expressing, Western Blot

9) Product Images from "miR-15a/16 inhibits TGF-beta3/VEGF signaling and increases retinal endothelial cell barrier proteins"

Article Title: miR-15a/16 inhibits TGF-beta3/VEGF signaling and increases retinal endothelial cell barrier proteins

Journal: Vision research

doi: 10.1016/j.visres.2017.07.007

Elevated levels of ZO-1 and occludin in REC. Western blot results for tight junction proteins on REC. Transfection with miR-15a, -16, and Neg. mimics were performed under HG conditions. HG decreased the levels of ZO-1 (A) and occludin (B) as compared to NG. REC transfected with miR-15a and/or -16 mimics (labeled as miR-15a, miR-16, and miR-15a/16) had increased levels of ZO-1 (A) and occludin (B), compared to that of control groups (HG and Neg.). # p
Figure Legend Snippet: Elevated levels of ZO-1 and occludin in REC. Western blot results for tight junction proteins on REC. Transfection with miR-15a, -16, and Neg. mimics were performed under HG conditions. HG decreased the levels of ZO-1 (A) and occludin (B) as compared to NG. REC transfected with miR-15a and/or -16 mimics (labeled as miR-15a, miR-16, and miR-15a/16) had increased levels of ZO-1 (A) and occludin (B), compared to that of control groups (HG and Neg.). # p

Techniques Used: Western Blot, Transfection, Labeling

Changes of TGF-beta3 signaling, ZO-1, and occludin levels in the retina of miR-15a overexpressing TG mice. Retinal lysates were examined by Western blot. The retinas of miR-15a-TG mice showed decreased levels of TGF- beta3 (A) and SMAD2/3 phosphoryation (B), with elevated levels of ZO-1 (C) and occludin (D) compared to control wild type (WT) mice. * p
Figure Legend Snippet: Changes of TGF-beta3 signaling, ZO-1, and occludin levels in the retina of miR-15a overexpressing TG mice. Retinal lysates were examined by Western blot. The retinas of miR-15a-TG mice showed decreased levels of TGF- beta3 (A) and SMAD2/3 phosphoryation (B), with elevated levels of ZO-1 (C) and occludin (D) compared to control wild type (WT) mice. * p

Techniques Used: Mouse Assay, Western Blot

10) Product Images from "Toll-Like Receptor 7 Agonist–Induced Dermatitis Causes Severe Dextran Sulfate Sodium Colitis by Altering the Gut Microbiome and Immune Cells"

Article Title: Toll-Like Receptor 7 Agonist–Induced Dermatitis Causes Severe Dextran Sulfate Sodium Colitis by Altering the Gut Microbiome and Immune Cells

Journal: Cellular and Molecular Gastroenterology and Hepatology

doi: 10.1016/j.jcmgh.2018.09.010

IMQ dermatitis mice did not increase intestinal permeability but altered immune cell composition in the intestine. ( A ) Analysis for intestinal permeability of mice induced psoriasis-like dermatitis (on day 17 of IMQ or vehicle treatment), by using 4 kilodaltons ( left ) or 70 kilodaltons ( right ) FITC-dextran. Plasma FITC-dextran concentrations were determined by fluorescence intensity, after 1, 2 and 4 hours from oral administration of FITC-dextran (n = 4, in each group). Seventy kilodaltons FITC-dextran was administered intravenously as a positive control. ( B ) Quantitative PCR analysis for ZO-1 and occludin in colonic epithelial cells after topical IMQ treatment (on day 17) analyzed by real-time PCR. ( C ) Western blot analysis for occludin expression in colonic epithelial cells after topical IMQ treatment (on day 17). Western blot ( left ) and relative protein levels ( right ) of occludin. ( D ) Immunofluorescence staining was performed in control and IMQ mice. ZO-1 ( top , green), occludin ( middle , red), claudin ( bottom , green) stained with 4′,6-diamidino-2-phenylindole (DAPI; blue) in colon tissue. Four to 5 sections from each mice were obtained and the average positive area of each protein was compared with the DAPI-positive area measured (n = 4–5, each dot represents each mouse). ( F ) Quantitative PCR analysis for Reg IIIγ in colonic epithelial cells after topical IMQ treatment (on day 17) analyzed by real-time PCR. ( G ) Representative plots of flow cytometry analysis for macrophages (CD11b + CD64 + cells) in CD45 + CD3 - CD19 - B220 - NK1.1 - colonic lamina propria after IMQ treatment (on day 17) (n = 6 in each group). Percentage ( left ) and absolute number ( right ) of macrophages in colonic lamina propria. ( H and I ) Percentage and absolute number of CD206 + ( left ) or CD80 + ( right ) macrophages. ( I ) Quantitative PCR analysis of IL10 , Tnf-α , Il6 , IL12b , Arg-1 , and Nos2 in LP CD11b + s (n = 3–4 in each group). ( J ) CD11b + cells were sorted by magnetic activated cell sorting from colonic lamina propria mononuclear cells on day 17 of IMQ treatment (LP CD11b + s). LP CD11b + s were cultured with lipopolysaccharide (LPS) or peptidoglycan (PGN) for 24 hours in vitro, and concentrations of cytokines in the supernatant were measured by cytometric bead array. Unstimulated control was cultured with medium (Med). ( K ) Western blot analysis for NLRP3 in peritoneal cavity cells, which are composed mainly of macrophages. Western blot ( left ) and relative protein levels ( right ) of NLRP3 (n = 7–8, pooled from 2 independent experiments, in each group). ( L ) Percentage of ILC3 in CD45 + CD3 - CD19 - cells and ( M ) pDCs in CD45 + CD11b - CD11c + cells in colonic lamina propria (ILC3, CD45 + CD3 - CD19 - Rorγt + cells; pDC, CD45 + CD11b - CD11c + B220 + PDCA-1 + cells. Each symbol represents an individual mouse (n = 4–8). Statistical analyses were performed with the Student t test. * P
Figure Legend Snippet: IMQ dermatitis mice did not increase intestinal permeability but altered immune cell composition in the intestine. ( A ) Analysis for intestinal permeability of mice induced psoriasis-like dermatitis (on day 17 of IMQ or vehicle treatment), by using 4 kilodaltons ( left ) or 70 kilodaltons ( right ) FITC-dextran. Plasma FITC-dextran concentrations were determined by fluorescence intensity, after 1, 2 and 4 hours from oral administration of FITC-dextran (n = 4, in each group). Seventy kilodaltons FITC-dextran was administered intravenously as a positive control. ( B ) Quantitative PCR analysis for ZO-1 and occludin in colonic epithelial cells after topical IMQ treatment (on day 17) analyzed by real-time PCR. ( C ) Western blot analysis for occludin expression in colonic epithelial cells after topical IMQ treatment (on day 17). Western blot ( left ) and relative protein levels ( right ) of occludin. ( D ) Immunofluorescence staining was performed in control and IMQ mice. ZO-1 ( top , green), occludin ( middle , red), claudin ( bottom , green) stained with 4′,6-diamidino-2-phenylindole (DAPI; blue) in colon tissue. Four to 5 sections from each mice were obtained and the average positive area of each protein was compared with the DAPI-positive area measured (n = 4–5, each dot represents each mouse). ( F ) Quantitative PCR analysis for Reg IIIγ in colonic epithelial cells after topical IMQ treatment (on day 17) analyzed by real-time PCR. ( G ) Representative plots of flow cytometry analysis for macrophages (CD11b + CD64 + cells) in CD45 + CD3 - CD19 - B220 - NK1.1 - colonic lamina propria after IMQ treatment (on day 17) (n = 6 in each group). Percentage ( left ) and absolute number ( right ) of macrophages in colonic lamina propria. ( H and I ) Percentage and absolute number of CD206 + ( left ) or CD80 + ( right ) macrophages. ( I ) Quantitative PCR analysis of IL10 , Tnf-α , Il6 , IL12b , Arg-1 , and Nos2 in LP CD11b + s (n = 3–4 in each group). ( J ) CD11b + cells were sorted by magnetic activated cell sorting from colonic lamina propria mononuclear cells on day 17 of IMQ treatment (LP CD11b + s). LP CD11b + s were cultured with lipopolysaccharide (LPS) or peptidoglycan (PGN) for 24 hours in vitro, and concentrations of cytokines in the supernatant were measured by cytometric bead array. Unstimulated control was cultured with medium (Med). ( K ) Western blot analysis for NLRP3 in peritoneal cavity cells, which are composed mainly of macrophages. Western blot ( left ) and relative protein levels ( right ) of NLRP3 (n = 7–8, pooled from 2 independent experiments, in each group). ( L ) Percentage of ILC3 in CD45 + CD3 - CD19 - cells and ( M ) pDCs in CD45 + CD11b - CD11c + cells in colonic lamina propria (ILC3, CD45 + CD3 - CD19 - Rorγt + cells; pDC, CD45 + CD11b - CD11c + B220 + PDCA-1 + cells. Each symbol represents an individual mouse (n = 4–8). Statistical analyses were performed with the Student t test. * P

Techniques Used: Mouse Assay, Permeability, Fluorescence, Positive Control, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence, Staining, Flow Cytometry, Cytometry, FACS, Cell Culture, In Vitro

11) Product Images from "Protective effects of lycium barbarum polysaccharides on blood-retinal barrier via ROCK1 pathway in diabetic rats"

Article Title: Protective effects of lycium barbarum polysaccharides on blood-retinal barrier via ROCK1 pathway in diabetic rats

Journal: American Journal of Translational Research

doi:

Western blot assay for P-Occludin, ROCK1 and P-MLC protein expression. The Western blot was performed at 4, 8 and 12 weeks after administration. The relative expression of P-Occludin, ROCK1 and P-MLC was displayed in (A-C) respectively. (D) Western blot assay. *P
Figure Legend Snippet: Western blot assay for P-Occludin, ROCK1 and P-MLC protein expression. The Western blot was performed at 4, 8 and 12 weeks after administration. The relative expression of P-Occludin, ROCK1 and P-MLC was displayed in (A-C) respectively. (D) Western blot assay. *P

Techniques Used: Western Blot, Expressing

Immunohistochemistry (IHC) assay for P-Occludin and ROCK1 protein expression. The IHC assay was performed at 4, 8 and 12 weeks after administration. CON: control group treated with normal saline; DM: model group treated with normal saline; LBP: model group treated with LBP; F: model group treated with fasudil; F + LBP: model group treated with fasudil and LBP.
Figure Legend Snippet: Immunohistochemistry (IHC) assay for P-Occludin and ROCK1 protein expression. The IHC assay was performed at 4, 8 and 12 weeks after administration. CON: control group treated with normal saline; DM: model group treated with normal saline; LBP: model group treated with LBP; F: model group treated with fasudil; F + LBP: model group treated with fasudil and LBP.

Techniques Used: Immunohistochemistry, Expressing

Western blot assay for P-Occludin, ROCK1 and P-MLC protein expression. The expression of Occudin, ROCK1 and P-MLC protein was evaluated by Western blot after 7 day incubation. A-C: Displayed the expression profile of P-Occludin, ROCK1 and P-MLC respectively. D: The photograph of Western blot band. *P
Figure Legend Snippet: Western blot assay for P-Occludin, ROCK1 and P-MLC protein expression. The expression of Occudin, ROCK1 and P-MLC protein was evaluated by Western blot after 7 day incubation. A-C: Displayed the expression profile of P-Occludin, ROCK1 and P-MLC respectively. D: The photograph of Western blot band. *P

Techniques Used: Western Blot, Expressing, Incubation

12) Product Images from "Co-Administration of Cholesterol-Lowering Probiotics and Anthraquinone from Cassia obtusifolia L. Ameliorate Non-Alcoholic Fatty Liver"

Article Title: Co-Administration of Cholesterol-Lowering Probiotics and Anthraquinone from Cassia obtusifolia L. Ameliorate Non-Alcoholic Fatty Liver

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138078

Probiotics and AC reduced endotoxin levels in rats and promoted ZO-1, occludin expression, and FXR mRNA expression. (A) Pathological morphology observed in rat ileum, Original magnification, ×100 (B) Serum levels of endotoxin. Data are expressed as mean±SD(n = 6). (C) Intestinal mRNA expression levels of the FXR genes. Results are expressed as mean±SD (n = 6) (D) Expression of ZO-1 and occludin were analysed by western bloting. The ß-actin expression was used as a loading control. Data are expressed as mean±SD (n = 3). All mean values within treatment groups with different lowercase letters are significantly different (P
Figure Legend Snippet: Probiotics and AC reduced endotoxin levels in rats and promoted ZO-1, occludin expression, and FXR mRNA expression. (A) Pathological morphology observed in rat ileum, Original magnification, ×100 (B) Serum levels of endotoxin. Data are expressed as mean±SD(n = 6). (C) Intestinal mRNA expression levels of the FXR genes. Results are expressed as mean±SD (n = 6) (D) Expression of ZO-1 and occludin were analysed by western bloting. The ß-actin expression was used as a loading control. Data are expressed as mean±SD (n = 3). All mean values within treatment groups with different lowercase letters are significantly different (P

Techniques Used: Expressing, Western Blot

13) Product Images from "Formononetin Administration Ameliorates Dextran Sulfate Sodium-Induced Acute Colitis by Inhibiting NLRP3 Inflammasome Signaling Pathway"

Article Title: Formononetin Administration Ameliorates Dextran Sulfate Sodium-Induced Acute Colitis by Inhibiting NLRP3 Inflammasome Signaling Pathway

Journal: Mediators of Inflammation

doi: 10.1155/2018/3048532

For protected against colonic epithelial tight junction injury and inhibited NLRP3 pathway in vitro. (a, b) Protein levels of NLRP3, ASC, and IL-1 β and (c, d) claudin-1, occludin, and ZO-1 were analyzed by western blotting. ∗ p
Figure Legend Snippet: For protected against colonic epithelial tight junction injury and inhibited NLRP3 pathway in vitro. (a, b) Protein levels of NLRP3, ASC, and IL-1 β and (c, d) claudin-1, occludin, and ZO-1 were analyzed by western blotting. ∗ p

Techniques Used: In Vitro, Western Blot

For relieved DSS-induced colonic epithelial tight junction destruction in mice. (a) Representative immunofluorescence images for claudin-1 (Green), occludin (Green), and ZO-1 (Green) in the colonic tissue. (b, c) Protein levels of claudin-1, occludin, and ZO-1 in the colon tissues were analyzed by western blotting. ∗∗ p
Figure Legend Snippet: For relieved DSS-induced colonic epithelial tight junction destruction in mice. (a) Representative immunofluorescence images for claudin-1 (Green), occludin (Green), and ZO-1 (Green) in the colonic tissue. (b, c) Protein levels of claudin-1, occludin, and ZO-1 in the colon tissues were analyzed by western blotting. ∗∗ p

Techniques Used: Mouse Assay, Immunofluorescence, Western Blot

NLRP3 inhibitor MCC950 eliminated the protective effect of H-For on acute colitis in mice. (a) The experimental protocol with For and MCC950 in acute colitis model. (b) Body weights of mice and (c) disease activity index (DAI) during the disease process. (d) Morphological changes in the mice colons, (e) variations of colon length of mice, (f) representative HE staining, and (g) histological scores of colonic tissue. (h) Protein levels of claudin-1, occludin, and ZO-1 and (i) NLRP3, ASC, and IL-1 β were analyzed by western blotting. ∗ p
Figure Legend Snippet: NLRP3 inhibitor MCC950 eliminated the protective effect of H-For on acute colitis in mice. (a) The experimental protocol with For and MCC950 in acute colitis model. (b) Body weights of mice and (c) disease activity index (DAI) during the disease process. (d) Morphological changes in the mice colons, (e) variations of colon length of mice, (f) representative HE staining, and (g) histological scores of colonic tissue. (h) Protein levels of claudin-1, occludin, and ZO-1 and (i) NLRP3, ASC, and IL-1 β were analyzed by western blotting. ∗ p

Techniques Used: Mouse Assay, Activity Assay, Staining, Western Blot

14) Product Images from "Hypercapnia exacerbates the disruption of the blood‑brain barrier by inducing interleukin‑1β overproduction in the blood of hypoxemic adult rats"

Article Title: Hypercapnia exacerbates the disruption of the blood‑brain barrier by inducing interleukin‑1β overproduction in the blood of hypoxemic adult rats

Journal: International Journal of Molecular Medicine

doi: 10.3892/ijmm.2020.4604

IL-1β treatment decreased the expression of tight junctional proteins in RBECs (n=4). (A) Immunoreactive bands of ZO-1 (220 kDa), occludin (59 kDa), claudin-5 (23 kDa) and β-actin (42 kDa). (B) There was decreased tight junctional protein expression in the IL-1β group compared with the control group (ZO-1: ** P
Figure Legend Snippet: IL-1β treatment decreased the expression of tight junctional proteins in RBECs (n=4). (A) Immunoreactive bands of ZO-1 (220 kDa), occludin (59 kDa), claudin-5 (23 kDa) and β-actin (42 kDa). (B) There was decreased tight junctional protein expression in the IL-1β group compared with the control group (ZO-1: ** P

Techniques Used: Expressing

Hypercapnia decreased the expression of tight junctional proteins in the hypoxemic hippocampus (n=4). (A) Immunoreactive bands of ZO-1 (220 kDa), occludin (59 kDa), claudin-5 (23 kDa) and β-actin (42 kDa). (B-D) There were interaction effects between hypoxia treatment and hypercapnia treatment (ZO-1: P
Figure Legend Snippet: Hypercapnia decreased the expression of tight junctional proteins in the hypoxemic hippocampus (n=4). (A) Immunoreactive bands of ZO-1 (220 kDa), occludin (59 kDa), claudin-5 (23 kDa) and β-actin (42 kDa). (B-D) There were interaction effects between hypoxia treatment and hypercapnia treatment (ZO-1: P

Techniques Used: Expressing

15) Product Images from "Tumor necrosis factor ligand-related molecule 1A affects the intestinal mucosal barrier function by promoting Th9/interleukin-9 expression"

Article Title: Tumor necrosis factor ligand-related molecule 1A affects the intestinal mucosal barrier function by promoting Th9/interleukin-9 expression

Journal: The Journal of International Medical Research

doi: 10.1177/0300060520926011

TL1A aggravates the intestinal mucosal barrier injury. (a) Immunofluorescence of occludin. (b) Immunofluorescence of claudin-1. (c, d) The expression of occludin, zonulin-1, and claudin-1 in Caco-2 cells. * P
Figure Legend Snippet: TL1A aggravates the intestinal mucosal barrier injury. (a) Immunofluorescence of occludin. (b) Immunofluorescence of claudin-1. (c, d) The expression of occludin, zonulin-1, and claudin-1 in Caco-2 cells. * P

Techniques Used: Immunofluorescence, Expressing

16) Product Images from "The Vacuolating Autotransporter Toxin (Vat) of Escherichia coli Causes Cell Cytoskeleton Changes and Produces Non-lysosomal Vacuole Formation in Bladder Epithelial Cells"

Article Title: The Vacuolating Autotransporter Toxin (Vat) of Escherichia coli Causes Cell Cytoskeleton Changes and Produces Non-lysosomal Vacuole Formation in Bladder Epithelial Cells

Journal: Frontiers in Cellular and Infection Microbiology

doi: 10.3389/fcimb.2020.00299

Modification of cell junctions of the bladder urothelial monolayer after incubation with Vat. Analysis by scanning electron microscopy revealed loss of integrity of the cell monolayer (White asterisks) starting at 6 h. The changes in the monolayer are more visible at 12 and 24 h after exposure to the toxin in comparison to the control. The immunofluorescence labeling of ZO-1 and Occludin proteins show cell junction disruption in the cell monolayer.
Figure Legend Snippet: Modification of cell junctions of the bladder urothelial monolayer after incubation with Vat. Analysis by scanning electron microscopy revealed loss of integrity of the cell monolayer (White asterisks) starting at 6 h. The changes in the monolayer are more visible at 12 and 24 h after exposure to the toxin in comparison to the control. The immunofluorescence labeling of ZO-1 and Occludin proteins show cell junction disruption in the cell monolayer.

Techniques Used: Modification, Incubation, Electron Microscopy, Immunofluorescence, Labeling

17) Product Images from "Cimifugin suppresses allergic inflammation by reducing epithelial derived initiative key factors via regulating tight junctions"

Article Title: Cimifugin suppresses allergic inflammation by reducing epithelial derived initiative key factors via regulating tight junctions

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13204

Effects of cimifugin on the TJ s expressions in the initial stage of AD model. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to GAPDH were quantified by ChemiScope analysis. ( E ), Immunohistochemical analysis of CLDN ‐1 and occludin expression ( n = 5, magnification: ×630). ( F–H ), CLDN ‐1 and occludin mean DAB intensity were quantified by Mantra Quantitative Pathology Workstation (mean + SD , n = 5, # P
Figure Legend Snippet: Effects of cimifugin on the TJ s expressions in the initial stage of AD model. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to GAPDH were quantified by ChemiScope analysis. ( E ), Immunohistochemical analysis of CLDN ‐1 and occludin expression ( n = 5, magnification: ×630). ( F–H ), CLDN ‐1 and occludin mean DAB intensity were quantified by Mantra Quantitative Pathology Workstation (mean + SD , n = 5, # P

Techniques Used: Western Blot, Immunohistochemistry, Expressing

Effects of cimifugin on CLDND 1, CLDN ‐1 and occludin in HaCaT cells. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot in HaCaT cells ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to β‐actin were quantified by ChemiScope analysis. ( E , F ), immunofluorescence quantification of CLDN ‐1 and occludin expressions ( n = 3, magnification: ×200; mean + SD , n = 3, # P
Figure Legend Snippet: Effects of cimifugin on CLDND 1, CLDN ‐1 and occludin in HaCaT cells. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot in HaCaT cells ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to β‐actin were quantified by ChemiScope analysis. ( E , F ), immunofluorescence quantification of CLDN ‐1 and occludin expressions ( n = 3, magnification: ×200; mean + SD , n = 3, # P

Techniques Used: Western Blot, Immunofluorescence

18) Product Images from "Cimifugin suppresses allergic inflammation by reducing epithelial derived initiative key factors via regulating tight junctions"

Article Title: Cimifugin suppresses allergic inflammation by reducing epithelial derived initiative key factors via regulating tight junctions

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13204

Effects of cimifugin on the TJ s expressions in the initial stage of AD model. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to GAPDH were quantified by ChemiScope analysis. ( E ), Immunohistochemical analysis of CLDN ‐1 and occludin expression ( n = 5, magnification: ×630). ( F–H ), CLDN ‐1 and occludin mean DAB intensity were quantified by Mantra Quantitative Pathology Workstation (mean + SD , n = 5, # P
Figure Legend Snippet: Effects of cimifugin on the TJ s expressions in the initial stage of AD model. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to GAPDH were quantified by ChemiScope analysis. ( E ), Immunohistochemical analysis of CLDN ‐1 and occludin expression ( n = 5, magnification: ×630). ( F–H ), CLDN ‐1 and occludin mean DAB intensity were quantified by Mantra Quantitative Pathology Workstation (mean + SD , n = 5, # P

Techniques Used: Western Blot, Immunohistochemistry, Expressing

Effects of cimifugin on CLDND 1, CLDN ‐1 and occludin in HaCaT cells. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot in HaCaT cells ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to β‐actin were quantified by ChemiScope analysis. ( E , F ), immunofluorescence quantification of CLDN ‐1 and occludin expressions ( n = 3, magnification: ×200; mean + SD , n = 3, # P
Figure Legend Snippet: Effects of cimifugin on CLDND 1, CLDN ‐1 and occludin in HaCaT cells. ( A ), CLDND 1, CLDN ‐1 and occludin expressions were analysed by Western blot in HaCaT cells ( n = 3). ( B–D ), CLDND 1, CLDN ‐1 and occludin expressions relative to β‐actin were quantified by ChemiScope analysis. ( E , F ), immunofluorescence quantification of CLDN ‐1 and occludin expressions ( n = 3, magnification: ×200; mean + SD , n = 3, # P

Techniques Used: Western Blot, Immunofluorescence

19) Product Images from "Mechanical Injury Induces Brain Endothelial-Derived Microvesicle Release: Implications for Cerebral Vascular Injury during Traumatic Brain Injury"

Article Title: Mechanical Injury Induces Brain Endothelial-Derived Microvesicle Release: Implications for Cerebral Vascular Injury during Traumatic Brain Injury

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2016.00043

In vitro mechanical injury induces the release of eMVs containing the endothelial marker PECAM-1 and tight junction protein occludin. (A) Injury increases the release of eMVs containing PECAM-1. eMVs were isolated from media using Exoquick and analyzed by western blot. Elevated levels of PECAM-1 were detected at 2 h and remained elevated for most time points. (Two-way ANOVA with Bonferroni post-hoc analysis ** p
Figure Legend Snippet: In vitro mechanical injury induces the release of eMVs containing the endothelial marker PECAM-1 and tight junction protein occludin. (A) Injury increases the release of eMVs containing PECAM-1. eMVs were isolated from media using Exoquick and analyzed by western blot. Elevated levels of PECAM-1 were detected at 2 h and remained elevated for most time points. (Two-way ANOVA with Bonferroni post-hoc analysis ** p

Techniques Used: In Vitro, Marker, Isolation, Western Blot

Experimental TBI induces the release of eMVs containing the tight junction protein occludin . Mice were subjected to sham surgery or a Controlled Cortical Impact, CCI-TBI and blood was collected 24 h post-injury. (A) Flow Cytometry detection of occludin (FITC) positive eMVs from blood plasma. Regions of interest were drawn around all FITC positive vesicles as well as a clear high FSC population. (B,C) TBI induces an increase in total FITC positive and high FSC-FITC positive vesicles at 24 h post-injury (Students two-tailed t -test average ± SEM n = 5 for sham and n = 4 for TBI, * p
Figure Legend Snippet: Experimental TBI induces the release of eMVs containing the tight junction protein occludin . Mice were subjected to sham surgery or a Controlled Cortical Impact, CCI-TBI and blood was collected 24 h post-injury. (A) Flow Cytometry detection of occludin (FITC) positive eMVs from blood plasma. Regions of interest were drawn around all FITC positive vesicles as well as a clear high FSC population. (B,C) TBI induces an increase in total FITC positive and high FSC-FITC positive vesicles at 24 h post-injury (Students two-tailed t -test average ± SEM n = 5 for sham and n = 4 for TBI, * p

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Two Tailed Test

20) Product Images from "Proteomic and cellular localisation studies suggest non‐tight junction cytoplasmic and nuclear roles for occludin in astrocytes, et al. Proteomic and cellular localisation studies suggest non‐tight junction cytoplasmic and nuclear roles for occludin in astrocytes"

Article Title: Proteomic and cellular localisation studies suggest non‐tight junction cytoplasmic and nuclear roles for occludin in astrocytes, et al. Proteomic and cellular localisation studies suggest non‐tight junction cytoplasmic and nuclear roles for occludin in astrocytes

Journal: The European Journal of Neuroscience

doi: 10.1111/ejn.13933

Immunohistochemistry for occludin in human autopsy brain. (a) Occludin pyramidal neuron staining with pyramidal nuclear reactivity indicated (arrows), score 2 (Case 14, MCI). (b) Cytoplasmic astrocyte expression (arrows, Case 21, AD). (c) Occludin reactivity around a plaque (arrows, Case 29, AD). (d) Occludin up‐regulation around ischaemic focus, inset showing up‐regulation in astrocytes (Case 15, MCI). (e) Occludin immunopositive glial nucleus (arrow). Note several adjacent negative nuclei. An occludin‐labelled capillary is also seen (arrowhead) (Case 11, MCI) (f) Dual labelling immunohistochemistry for occludin (brown) and GFAP (pink). Arrows show occludin positive GFAP‐astrocytes [Colour figure can be viewed at http://wileyonlinelibrary.com ]
Figure Legend Snippet: Immunohistochemistry for occludin in human autopsy brain. (a) Occludin pyramidal neuron staining with pyramidal nuclear reactivity indicated (arrows), score 2 (Case 14, MCI). (b) Cytoplasmic astrocyte expression (arrows, Case 21, AD). (c) Occludin reactivity around a plaque (arrows, Case 29, AD). (d) Occludin up‐regulation around ischaemic focus, inset showing up‐regulation in astrocytes (Case 15, MCI). (e) Occludin immunopositive glial nucleus (arrow). Note several adjacent negative nuclei. An occludin‐labelled capillary is also seen (arrowhead) (Case 11, MCI) (f) Dual labelling immunohistochemistry for occludin (brown) and GFAP (pink). Arrows show occludin positive GFAP‐astrocytes [Colour figure can be viewed at http://wileyonlinelibrary.com ]

Techniques Used: Immunohistochemistry, Staining, Expressing

Occludin expression in human primary astrocytes and the human astrocytoma line 1321N1. (a) Western blotting confirms expression of the protein at the expected size as well as additional bands (see main text for description), molecular weight markers are indicated in kDa. (b) Immunocytochemistry for occludin (red) demonstrates both cytoplasmic and nuclear expression of the protein. Nuclei are shown in blue (Hoescht), scale bar represents 10μm [Colour figure can be viewed at http://wileyonlinelibrary.com ]
Figure Legend Snippet: Occludin expression in human primary astrocytes and the human astrocytoma line 1321N1. (a) Western blotting confirms expression of the protein at the expected size as well as additional bands (see main text for description), molecular weight markers are indicated in kDa. (b) Immunocytochemistry for occludin (red) demonstrates both cytoplasmic and nuclear expression of the protein. Nuclei are shown in blue (Hoescht), scale bar represents 10μm [Colour figure can be viewed at http://wileyonlinelibrary.com ]

Techniques Used: Expressing, Western Blot, Molecular Weight, Immunocytochemistry

Occludin expression in fractionated human 1321N cells. Western blotting using an antibody directed against the C‐terminal of occludin (a) and the N‐terminal of occludin (b) confirms expression at the expected molecular weights. Whole cell lysates from the endothelial cell line hCMEC/D3 were included as a positive control. Cell fractionation was confirmed using an antibody directed against SSRP1 (nucleus) and HSPA14 (cytoplasm). Molecular weight markers are indicated in kDa
Figure Legend Snippet: Occludin expression in fractionated human 1321N cells. Western blotting using an antibody directed against the C‐terminal of occludin (a) and the N‐terminal of occludin (b) confirms expression at the expected molecular weights. Whole cell lysates from the endothelial cell line hCMEC/D3 were included as a positive control. Cell fractionation was confirmed using an antibody directed against SSRP1 (nucleus) and HSPA14 (cytoplasm). Molecular weight markers are indicated in kDa

Techniques Used: Expressing, Western Blot, Positive Control, Cell Fractionation, Molecular Weight

21) Product Images from "Hydrogen Sulfide Ameliorates Blood-Spinal Cord Barrier Disruption and Improves Functional Recovery by Inhibiting Endoplasmic Reticulum Stress-Dependent Autophagy"

Article Title: Hydrogen Sulfide Ameliorates Blood-Spinal Cord Barrier Disruption and Improves Functional Recovery by Inhibiting Endoplasmic Reticulum Stress-Dependent Autophagy

Journal: Frontiers in Pharmacology

doi: 10.3389/fphar.2018.00858

NaHS protects OGD-treated HUVECs by inhibiting ER stress and autophagy in vitro . (A) Representative western blots of P120, β-catenin and Occludin for each group. (B–D) Quantification of the expression of P120, β-catenin and Occludin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM. ∗∗∗ P
Figure Legend Snippet: NaHS protects OGD-treated HUVECs by inhibiting ER stress and autophagy in vitro . (A) Representative western blots of P120, β-catenin and Occludin for each group. (B–D) Quantification of the expression of P120, β-catenin and Occludin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM. ∗∗∗ P

Techniques Used: In Vitro, Western Blot, Expressing

NaHS prevents the loss of TJ and AJ proteins by inhibiting ER stress and autophagy at 1d post-SCI. (A) TM and Rapa were applied to specifically activate ER stress and autophagy, respectively. The rats were randomly divided into five groups: (i) the Sham group; (ii) the SCI model group; (iii) the SCI model with NaHS treatment (SCI + NaHS) group; (iv) the SCI model with NaHS and TM treatment (SCI + NaHS + TM) group; and (v) the SCI model with NaHS and Rapa treatment (SCI + NaHS + Rapa) group. Representative western blots of P120, β-catenin and Occludin from each group. (B–D) Quantification of the expression of P120, β-catenin, and Occludin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM, n = 5. ∗∗∗ P
Figure Legend Snippet: NaHS prevents the loss of TJ and AJ proteins by inhibiting ER stress and autophagy at 1d post-SCI. (A) TM and Rapa were applied to specifically activate ER stress and autophagy, respectively. The rats were randomly divided into five groups: (i) the Sham group; (ii) the SCI model group; (iii) the SCI model with NaHS treatment (SCI + NaHS) group; (iv) the SCI model with NaHS and TM treatment (SCI + NaHS + TM) group; and (v) the SCI model with NaHS and Rapa treatment (SCI + NaHS + Rapa) group. Representative western blots of P120, β-catenin and Occludin from each group. (B–D) Quantification of the expression of P120, β-catenin, and Occludin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM, n = 5. ∗∗∗ P

Techniques Used: Western Blot, Expressing

SCI induces the loss of TJ and AJ proteins. (A) Representative western blotting results for TJ proteins (Occludin) and AJ proteins (P120 and β-catenin) in the sham group, 6 h, 12 h, 1d, 3d, and 7d after SCI group. (B–D) Quantification of the expression of Occludin, P120, and β-catenin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM, n = 5. ∗ P
Figure Legend Snippet: SCI induces the loss of TJ and AJ proteins. (A) Representative western blotting results for TJ proteins (Occludin) and AJ proteins (P120 and β-catenin) in the sham group, 6 h, 12 h, 1d, 3d, and 7d after SCI group. (B–D) Quantification of the expression of Occludin, P120, and β-catenin. The relative band density value was normalized to that of GAPDH. All data are presented as the mean ± SEM, n = 5. ∗ P

Techniques Used: Western Blot, Expressing

22) Product Images from "Replacement of grains with soybean hulls ameliorates SARA-induced impairment of the colonic epithelium barrier function of goats"

Article Title: Replacement of grains with soybean hulls ameliorates SARA-induced impairment of the colonic epithelium barrier function of goats

Journal: BMC Veterinary Research

doi: 10.1186/s12917-018-1705-8

Protein abundance of claudin-1, occludin, and ZO-1 in the colonic epithelium of goats in the LC, HC, and SH groups. The abundance of claudin-1 and ZO-1 proteins in the HC group significantly decreased compared with that in the LC group, while the levels in the SH group were not significantly altered. The results are expressed as fold changes relative to those in the LC group (mean ± SEM). * indicates P
Figure Legend Snippet: Protein abundance of claudin-1, occludin, and ZO-1 in the colonic epithelium of goats in the LC, HC, and SH groups. The abundance of claudin-1 and ZO-1 proteins in the HC group significantly decreased compared with that in the LC group, while the levels in the SH group were not significantly altered. The results are expressed as fold changes relative to those in the LC group (mean ± SEM). * indicates P

Techniques Used:

The expression of genes related to tight junction proteins in the colonic epithelium of goats in the LC, HC, and SH groups. The colonic epithelium of the goat fed the HC diet had a significant decline in the mRNA expression of claudin-1, claudin-4, occludin, and ZO-1 compared with that in the LC group, while no significant change between the SH and LC groups was observed. The results are expressed as fold changes relative to those in the LC group (mean ± SEM). * indicates P
Figure Legend Snippet: The expression of genes related to tight junction proteins in the colonic epithelium of goats in the LC, HC, and SH groups. The colonic epithelium of the goat fed the HC diet had a significant decline in the mRNA expression of claudin-1, claudin-4, occludin, and ZO-1 compared with that in the LC group, while no significant change between the SH and LC groups was observed. The results are expressed as fold changes relative to those in the LC group (mean ± SEM). * indicates P

Techniques Used: Expressing

23) Product Images from "Structural alterations of the intestinal epithelial barrier in Parkinson’s disease"

Article Title: Structural alterations of the intestinal epithelial barrier in Parkinson’s disease

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-015-0196-0

Expression of TJs proteins in colonic biopsies from patients with Parkinson’s disease (PD) and control subjects (CTRL). Biopsies lysates (20 μg of protein per sample) were subjected to immunoblot analysis using antibodies against occludin and ZO-1 (A) . Beta-actin was used as a loading control. The optical densities of occludin (B) and ZO-1 (C) immunoreactive bands were measured, normalized to the optical densities of beta-actin immunoreactive bands in the same samples and expressed as percentages of controls. Data correspond to mean ± SEM of 11 samples for control subjects (CTRL) and 31 samples for Parkinson’s disease (PD) patients. Patients versus control, *: p
Figure Legend Snippet: Expression of TJs proteins in colonic biopsies from patients with Parkinson’s disease (PD) and control subjects (CTRL). Biopsies lysates (20 μg of protein per sample) were subjected to immunoblot analysis using antibodies against occludin and ZO-1 (A) . Beta-actin was used as a loading control. The optical densities of occludin (B) and ZO-1 (C) immunoreactive bands were measured, normalized to the optical densities of beta-actin immunoreactive bands in the same samples and expressed as percentages of controls. Data correspond to mean ± SEM of 11 samples for control subjects (CTRL) and 31 samples for Parkinson’s disease (PD) patients. Patients versus control, *: p

Techniques Used: Expressing

Localization of TJs proteins in the colonic mucosa of healthy controls (CTRL) and patients with Parkinson’s disease (PD). Representative photomicrographs of the colonic mucosa labeled with antibodies against ZO-1 (A, B) and occludin (C, D) in the colonic mucosa of control and PD patients; scale bar: 100 μm. High-magnification image of each area marked by red square; scale bar: 10 μm.
Figure Legend Snippet: Localization of TJs proteins in the colonic mucosa of healthy controls (CTRL) and patients with Parkinson’s disease (PD). Representative photomicrographs of the colonic mucosa labeled with antibodies against ZO-1 (A, B) and occludin (C, D) in the colonic mucosa of control and PD patients; scale bar: 100 μm. High-magnification image of each area marked by red square; scale bar: 10 μm.

Techniques Used: Labeling

24) Product Images from "Human iPS derived progenitors bioengineered into liver organoids using an inverted colloidal crystal poly (ethylene glycol) scaffold"

Article Title: Human iPS derived progenitors bioengineered into liver organoids using an inverted colloidal crystal poly (ethylene glycol) scaffold

Journal: Biomaterials

doi: 10.1016/j.biomaterials.2018.07.043

Disease modelling and in vivo transplantation. ( A ) Heatmap and hierarchal clustering comparing expression of 12 genes involved in encoding HCV entry and assembly in IH-ICC vs 2D vs primary (adult, fetal) liver. ( B ) Confocal imaging showing expression of claudin 1 and occludin in IH-ICC organoids. Scale bar, 100 μm. White and red arrowheads point to apical and lateral regions respectively. ( C ) HCV expression of IH-ICC vs 2D following infection with HCV reporter virus expressing secreted GLuc (HCVcc, N = 4) or mock infected with knock down HCVcc (kd-HCVcc, N = 3) and subsequently were sampled and washed daily. RLU, relative luminescence unit. ( D ) Photograph showing location of surgical pocket formation on murine left lateral lobe (left) and appearance following IH-ICC transplantation (right). The white dashed line depicts the capsular incision and the limits of the sub-capsular scaffold implant are shown by the white arrows. Scale bar 1.5 mm ( E ) H E staining of explant reveals neo-vasculature of IH-ICC. Scale bar, 100 μm. ( F ) Immuno-histochemical staining of explant for human albumin. Dashed white line indicates the boundary between implant and host liver. Scale bar, 100 μm. Mean ± sd; **p
Figure Legend Snippet: Disease modelling and in vivo transplantation. ( A ) Heatmap and hierarchal clustering comparing expression of 12 genes involved in encoding HCV entry and assembly in IH-ICC vs 2D vs primary (adult, fetal) liver. ( B ) Confocal imaging showing expression of claudin 1 and occludin in IH-ICC organoids. Scale bar, 100 μm. White and red arrowheads point to apical and lateral regions respectively. ( C ) HCV expression of IH-ICC vs 2D following infection with HCV reporter virus expressing secreted GLuc (HCVcc, N = 4) or mock infected with knock down HCVcc (kd-HCVcc, N = 3) and subsequently were sampled and washed daily. RLU, relative luminescence unit. ( D ) Photograph showing location of surgical pocket formation on murine left lateral lobe (left) and appearance following IH-ICC transplantation (right). The white dashed line depicts the capsular incision and the limits of the sub-capsular scaffold implant are shown by the white arrows. Scale bar 1.5 mm ( E ) H E staining of explant reveals neo-vasculature of IH-ICC. Scale bar, 100 μm. ( F ) Immuno-histochemical staining of explant for human albumin. Dashed white line indicates the boundary between implant and host liver. Scale bar, 100 μm. Mean ± sd; **p

Techniques Used: In Vivo, Transplantation Assay, Expressing, Immunocytochemistry, Imaging, Infection, Staining

25) Product Images from "Donor cornea transfer from Optisol GS to organ culture storage: a two-step procedure to increase donor tissue lifespan"

Article Title: Donor cornea transfer from Optisol GS to organ culture storage: a two-step procedure to increase donor tissue lifespan

Journal: Acta Ophthalmologica

doi: 10.1111/j.1755-3768.2012.02390.x

Real-time RT-PCR analysis for p63 , Connexin-43 , Ki-67 , Occludin , E-Cadherin, ABCG2 and P53 genes.
Figure Legend Snippet: Real-time RT-PCR analysis for p63 , Connexin-43 , Ki-67 , Occludin , E-Cadherin, ABCG2 and P53 genes.

Techniques Used: Quantitative RT-PCR

Immunostaining of adherens, tight and gap junction-associated markers Occludin (A, B), E-cadherin (C, D) and Connexin-43 (E, F) after storage in Optisol GS and after Optisol GS + EBOC. EBOC, Eye Bank Organ Culture.
Figure Legend Snippet: Immunostaining of adherens, tight and gap junction-associated markers Occludin (A, B), E-cadherin (C, D) and Connexin-43 (E, F) after storage in Optisol GS and after Optisol GS + EBOC. EBOC, Eye Bank Organ Culture.

Techniques Used: Immunostaining, Organ Culture

26) Product Images from "Caffeine protects against MPTP-induced blood-brain barrier dysfunction in mouse striatum"

Article Title: Caffeine protects against MPTP-induced blood-brain barrier dysfunction in mouse striatum

Journal:

doi: 10.1111/j.1471-4159.2008.05697.x

Caffeine blocked MPTP-induced decreases in occludin expression levels
Figure Legend Snippet: Caffeine blocked MPTP-induced decreases in occludin expression levels

Techniques Used: Expressing

27) Product Images from "An Intestinal Paracellular Pathway Biased Toward Positively-Charged Macromolecules"

Article Title: An Intestinal Paracellular Pathway Biased Toward Positively-Charged Macromolecules

Journal: Journal of controlled release : official journal of the Controlled Release Society

doi: 10.1016/j.jconrel.2018.09.003

PIP 640 fails to affect cellular levels or distribution of TAMPs (occludin, tricellulin, and marvelD3) as well as the scaffolding protein ZO-1 in Caco-2 cell monolayers in vitro. A) Representative immunoblots of TAMPS and ZO-1 total protein levels showed
Figure Legend Snippet: PIP 640 fails to affect cellular levels or distribution of TAMPs (occludin, tricellulin, and marvelD3) as well as the scaffolding protein ZO-1 in Caco-2 cell monolayers in vitro. A) Representative immunoblots of TAMPS and ZO-1 total protein levels showed

Techniques Used: Scaffolding, In Vitro, Western Blot

Actions of PIP 640 on Caco-2 cell monolayers are distinct from those of the pro-inflammatory cytokines TNF-α/INFγ. or trans-epithelial electrical resistance (TEER) and occludin protein levels. A) Caco-2 monolayer TEER value changes after
Figure Legend Snippet: Actions of PIP 640 on Caco-2 cell monolayers are distinct from those of the pro-inflammatory cytokines TNF-α/INFγ. or trans-epithelial electrical resistance (TEER) and occludin protein levels. A) Caco-2 monolayer TEER value changes after

Techniques Used:

28) Product Images from "FoxO1 regulates TLR4/MyD88/MD2‐NF‐κB inflammatory signalling in mucosal barrier injury of inflammatory bowel disease, et al. FoxO1 regulates TLR4/MyD88/MD2‐NF‐κB inflammatory signalling in mucosal barrier injury of inflammatory bowel disease"

Article Title: FoxO1 regulates TLR4/MyD88/MD2‐NF‐κB inflammatory signalling in mucosal barrier injury of inflammatory bowel disease, et al. FoxO1 regulates TLR4/MyD88/MD2‐NF‐κB inflammatory signalling in mucosal barrier injury of inflammatory bowel disease

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.15075

Effects of FoxO1 on the expression of tight junction protein in mice with chronic colitis. A, Expression of ZO‐1 protein in mouse intestinal tissue by IHC: ZO‐1 protein was strongly expressed in intestinal tissues of FoxO1‐WT‐Con and FoxO1‐Tg‐Con groups, which was significantly down‐regulated in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups. Meanwhile, the expression of ZO‐1 was significantly lower in FoxO1‐Tg‐DSS group than that of FoxO1‐WT‐DSS group. B, Expression of occludin protein in mouse intestinal tissue by IHC: Occludin protein was strongly expressed in intestinal tissues of FoxO1‐WT‐Con and FoxO1‐Tg‐Con groups, which was significantly down‐regulated in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups. Meanwhile, the expression of occludin was significantly lower in FoxO1‐Tg‐DSS group than that of FoxO1‐WT‐DSS group. C, Expression of FoxO1 protein in mouse intestinal tissue by IHC: The expression levels of FoxO1 were significantly increased in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups after DSS intervention, whereas the expression levels of FoxO1 were significantly higher in FoxO1‐Tg‐DSS group than those in FoxO1‐WT‐DSS group. D,E, The protein expression of ZO‐1 and occludin in intestinal tissue by Western blot. After DSS intervention, the expression levels of ZO‐1 and occludin were down‐regulated, whereas the expression levels of ZO‐1 and occludin were significantly lower in FoxO1‐Tg‐DSS than those in FoxO1‐WT‐DSS group. Comparison between groups, * P
Figure Legend Snippet: Effects of FoxO1 on the expression of tight junction protein in mice with chronic colitis. A, Expression of ZO‐1 protein in mouse intestinal tissue by IHC: ZO‐1 protein was strongly expressed in intestinal tissues of FoxO1‐WT‐Con and FoxO1‐Tg‐Con groups, which was significantly down‐regulated in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups. Meanwhile, the expression of ZO‐1 was significantly lower in FoxO1‐Tg‐DSS group than that of FoxO1‐WT‐DSS group. B, Expression of occludin protein in mouse intestinal tissue by IHC: Occludin protein was strongly expressed in intestinal tissues of FoxO1‐WT‐Con and FoxO1‐Tg‐Con groups, which was significantly down‐regulated in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups. Meanwhile, the expression of occludin was significantly lower in FoxO1‐Tg‐DSS group than that of FoxO1‐WT‐DSS group. C, Expression of FoxO1 protein in mouse intestinal tissue by IHC: The expression levels of FoxO1 were significantly increased in FoxO1‐WT‐DSS and FoxO1‐Tg‐DSS groups after DSS intervention, whereas the expression levels of FoxO1 were significantly higher in FoxO1‐Tg‐DSS group than those in FoxO1‐WT‐DSS group. D,E, The protein expression of ZO‐1 and occludin in intestinal tissue by Western blot. After DSS intervention, the expression levels of ZO‐1 and occludin were down‐regulated, whereas the expression levels of ZO‐1 and occludin were significantly lower in FoxO1‐Tg‐DSS than those in FoxO1‐WT‐DSS group. Comparison between groups, * P

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Western Blot

29) Product Images from "High-Glucose or -Fructose Diet Cause Changes of the Gut Microbiota and Metabolic Disorders in Mice without Body Weight Change"

Article Title: High-Glucose or -Fructose Diet Cause Changes of the Gut Microbiota and Metabolic Disorders in Mice without Body Weight Change

Journal: Nutrients

doi: 10.3390/nu10060761

HFD, HGD or HFrD-induced changes of gut permeability and related proteins expression. ( A ) plasma fluorescein isothiocyanate (FITC)-dextran concentration; ( B ) AUC of Plasma FITC-dextran levels; ( C ) representative images of Western blots for tight junction proteins (Occludin and ZO1) and inflammatory cytokines (IL-1β and TNF-α); ( D – G ) relative band intensities of Occludin ( D ), ZO1 ( E ), IL-1β ( F ) and TNF-α ( G ) normalized to those of β-actin. Data are presented as mean ± SEM for 9 mice per group ( A , B ) and mean percentage of ND ± SEM of three independent experiments (C-G) (* p
Figure Legend Snippet: HFD, HGD or HFrD-induced changes of gut permeability and related proteins expression. ( A ) plasma fluorescein isothiocyanate (FITC)-dextran concentration; ( B ) AUC of Plasma FITC-dextran levels; ( C ) representative images of Western blots for tight junction proteins (Occludin and ZO1) and inflammatory cytokines (IL-1β and TNF-α); ( D – G ) relative band intensities of Occludin ( D ), ZO1 ( E ), IL-1β ( F ) and TNF-α ( G ) normalized to those of β-actin. Data are presented as mean ± SEM for 9 mice per group ( A , B ) and mean percentage of ND ± SEM of three independent experiments (C-G) (* p

Techniques Used: Permeability, Expressing, Concentration Assay, Western Blot, Mouse Assay

30) Product Images from "Nitrosporeusine analogue ameliorates Chandipura virus induced inflammatory response in CNS via NFκb inactivation in microglia"

Article Title: Nitrosporeusine analogue ameliorates Chandipura virus induced inflammatory response in CNS via NFκb inactivation in microglia

Journal: PLoS Neglected Tropical Diseases

doi: 10.1371/journal.pntd.0006648

Leukocyte infiltration and BBB breaching during CHPV infection. A). Peripheral immune cells were stained to check their presence in brain post infection. CD3 positive cell were more prominent in CHPV infected samples. Similarly CD68 and CD11b positive cells were present in CHPV infected sample suggesting infiltration of peripheral cells in brain. B). To examine BBB breaching, we checked expression level of tight junction protein. Claudin-1, occludin and β-catenine was checked for western blot. Data suggest increase in expression of these proteins in CHPV infected samples. (-)-25b treated brain shows significant decrease in expression level of the proteins. C). mRNA expression was checked to analyse the status of ICAM, VCAM and MMP-9. Validation of results were done by 3 independent experiments with at least 4 animals in each group.*p
Figure Legend Snippet: Leukocyte infiltration and BBB breaching during CHPV infection. A). Peripheral immune cells were stained to check their presence in brain post infection. CD3 positive cell were more prominent in CHPV infected samples. Similarly CD68 and CD11b positive cells were present in CHPV infected sample suggesting infiltration of peripheral cells in brain. B). To examine BBB breaching, we checked expression level of tight junction protein. Claudin-1, occludin and β-catenine was checked for western blot. Data suggest increase in expression of these proteins in CHPV infected samples. (-)-25b treated brain shows significant decrease in expression level of the proteins. C). mRNA expression was checked to analyse the status of ICAM, VCAM and MMP-9. Validation of results were done by 3 independent experiments with at least 4 animals in each group.*p

Techniques Used: Infection, Staining, Expressing, Western Blot

31) Product Images from "A Possible Molecular Mechanism of Hearing loss during cerebral ischemia in mice"

Article Title: A Possible Molecular Mechanism of Hearing loss during cerebral ischemia in mice

Journal: Canadian journal of physiology and pharmacology

doi: 10.1139/cjpp-2014-0489

Western blot analysis of tight junction proteins Western blot and bar graph represents expression of Claudin-5 and Occludin in sham and ischemic group of animals (4A–B) . A *P
Figure Legend Snippet: Western blot analysis of tight junction proteins Western blot and bar graph represents expression of Claudin-5 and Occludin in sham and ischemic group of animals (4A–B) . A *P

Techniques Used: Western Blot, Expressing

32) Product Images from "Synergistic protection of astragalus polysaccharides and matrine against ulcerative colitis and associated lung injury in rats"

Article Title: Synergistic protection of astragalus polysaccharides and matrine against ulcerative colitis and associated lung injury in rats

Journal: World Journal of Gastroenterology

doi: 10.3748/wjg.v26.i1.55

Astragalus polysaccharides combined with matrine inhibit the expression of zonula occludens-1 and Occludin in lung and colon tissues of rats with ulcerative colitis. A and B: Zonula occludens-1 (ZO-1) expression in colon (A) and lung tissues (B) in various groups detected by Western blot analysis; C and D: Occludin expression in colon (C) and lung tissues (D) in various groups detected by immunohistochemistry analysis. a P
Figure Legend Snippet: Astragalus polysaccharides combined with matrine inhibit the expression of zonula occludens-1 and Occludin in lung and colon tissues of rats with ulcerative colitis. A and B: Zonula occludens-1 (ZO-1) expression in colon (A) and lung tissues (B) in various groups detected by Western blot analysis; C and D: Occludin expression in colon (C) and lung tissues (D) in various groups detected by immunohistochemistry analysis. a P

Techniques Used: Expressing, Western Blot, Immunohistochemistry

33) Product Images from "TGFβ-activated Kinase 1 (TAK1) Inhibition by 5Z-7-Oxozeaenol Attenuates Early Brain Injury after Experimental Subarachnoid Hemorrhage *"

Article Title: TGFβ-activated Kinase 1 (TAK1) Inhibition by 5Z-7-Oxozeaenol Attenuates Early Brain Injury after Experimental Subarachnoid Hemorrhage *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M115.636795

TAK1 inhibition failed to attenuate SAH-induced cerebral edema. OZ treatment did not reverse brain water content ( A ) and the levels of ZO-1 ( B ) and occludin ( C ) 24 h post-SAH. #, p
Figure Legend Snippet: TAK1 inhibition failed to attenuate SAH-induced cerebral edema. OZ treatment did not reverse brain water content ( A ) and the levels of ZO-1 ( B ) and occludin ( C ) 24 h post-SAH. #, p

Techniques Used: Inhibition

34) Product Images from "Activation of GABAA Receptors in Colon Epithelium Exacerbates Acute Colitis"

Article Title: Activation of GABAA Receptors in Colon Epithelium Exacerbates Acute Colitis

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2018.00987

Gamma-aminobutyric acid (GABA) reduces the expression of occludin and junctional adhesion molecule 1 (JAM-1) in mice. (A) Representative immunohistochemical data for occludin staining (brown) in colon mucosa within the different groups, scale bar 100 μm. (B) Representative immunohistochemical data for JAM-1 staining (brown) in colon mucosa within the different groups, scale bar 100 μm. (C) Representative immunofluorescence staining of JAM-1 (red) in DSS-treated Caco-2 cells with or without GABA, scale bar 25 μm.
Figure Legend Snippet: Gamma-aminobutyric acid (GABA) reduces the expression of occludin and junctional adhesion molecule 1 (JAM-1) in mice. (A) Representative immunohistochemical data for occludin staining (brown) in colon mucosa within the different groups, scale bar 100 μm. (B) Representative immunohistochemical data for JAM-1 staining (brown) in colon mucosa within the different groups, scale bar 100 μm. (C) Representative immunofluorescence staining of JAM-1 (red) in DSS-treated Caco-2 cells with or without GABA, scale bar 25 μm.

Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining, Immunofluorescence

35) Product Images from "Acupoint Catgut Embedding Improves the Lipopolysaccharide-Induced Acute Respiratory Distress Syndrome in Rats"

Article Title: Acupoint Catgut Embedding Improves the Lipopolysaccharide-Induced Acute Respiratory Distress Syndrome in Rats

Journal: BioMed Research International

doi: 10.1155/2020/2394734

ACE mitigates the LPS-downregulated AQP1 and Occludin proteins in the lung tissues. AQP1 (a), Occludin (b), and β -actin proteins in the lung tissues of each animal were quantified by Western blot. Data are representative images or expressed as the mean ± SD of individual groups ( n = 5 per group) from three separate experiments. AQP1: aquaporin 1. ∗ P
Figure Legend Snippet: ACE mitigates the LPS-downregulated AQP1 and Occludin proteins in the lung tissues. AQP1 (a), Occludin (b), and β -actin proteins in the lung tissues of each animal were quantified by Western blot. Data are representative images or expressed as the mean ± SD of individual groups ( n = 5 per group) from three separate experiments. AQP1: aquaporin 1. ∗ P

Techniques Used: Western Blot

36) Product Images from "Rottlerin ameliorates DSS-induced colitis by improving intestinal barrier function via activation of the Epac-2/Rap-1 signaling pathway"

Article Title: Rottlerin ameliorates DSS-induced colitis by improving intestinal barrier function via activation of the Epac-2/Rap-1 signaling pathway

Journal: bioRxiv

doi: 10.1101/2020.03.16.994582

Rottlerin treatment improved the distribution of TJ proteins and reduced the permeability of the intestine (A-B) IF revealed that the distributions of occludin and ZO-1 (green) in the colon of rottlerin group were dramatically upregulated in contrast to the DSS-treated group. The nuclei was stained by DAPI (blue). (C-D) This trend was confirmed again by the results of western blotting. (E) Intestinal permeability in mice was detected, the datas suggested that serum glucan binding levels in the rottlerin team were below those in the only DSS-treated mice, but similar to the WT mice. (F-G) In the rottlerin group, the probability of bacterial transfer to liver and MLN was below the DSS-treated group, but similar to the WT mice. The detection were executed 3 times separately (8 mice of every group), a representative result is given. The datas are appeared as the means ± SD (* P
Figure Legend Snippet: Rottlerin treatment improved the distribution of TJ proteins and reduced the permeability of the intestine (A-B) IF revealed that the distributions of occludin and ZO-1 (green) in the colon of rottlerin group were dramatically upregulated in contrast to the DSS-treated group. The nuclei was stained by DAPI (blue). (C-D) This trend was confirmed again by the results of western blotting. (E) Intestinal permeability in mice was detected, the datas suggested that serum glucan binding levels in the rottlerin team were below those in the only DSS-treated mice, but similar to the WT mice. (F-G) In the rottlerin group, the probability of bacterial transfer to liver and MLN was below the DSS-treated group, but similar to the WT mice. The detection were executed 3 times separately (8 mice of every group), a representative result is given. The datas are appeared as the means ± SD (* P

Techniques Used: Permeability, Staining, Western Blot, Mouse Assay, Binding Assay

Rottlerin upregulated the levels of TJ proteins by activating the Epac-2/Rap-1 pathway (A-B) IF revealed that the distributions of occludin and ZO-1 (green) was enhanced in the rottlerin group compared with the LPS - treated FHs 74 int cells but that HJC0197 inhibited this trend. The nuclei was stained by DAPI (blue). (C-D) The levels of occludin and ZO-1 in the rottlerin group were clearly raised in contrast to the LPS - treated FHs 74 int cells and were similar to those in the control group, but HJC0197 inhibited the effect of rottlerin. The detection were executed 3 times separately (8 mice of every group), a representative result is given. The datas are appeared as the means ± SD (* P
Figure Legend Snippet: Rottlerin upregulated the levels of TJ proteins by activating the Epac-2/Rap-1 pathway (A-B) IF revealed that the distributions of occludin and ZO-1 (green) was enhanced in the rottlerin group compared with the LPS - treated FHs 74 int cells but that HJC0197 inhibited this trend. The nuclei was stained by DAPI (blue). (C-D) The levels of occludin and ZO-1 in the rottlerin group were clearly raised in contrast to the LPS - treated FHs 74 int cells and were similar to those in the control group, but HJC0197 inhibited the effect of rottlerin. The detection were executed 3 times separately (8 mice of every group), a representative result is given. The datas are appeared as the means ± SD (* P

Techniques Used: Staining, Mouse Assay

37) Product Images from "Mfsd2a Reverses Spatial Learning and Memory Impairment Caused by Chronic Cerebral Hypoperfusion via Protection of the Blood–Brain Barrier"

Article Title: Mfsd2a Reverses Spatial Learning and Memory Impairment Caused by Chronic Cerebral Hypoperfusion via Protection of the Blood–Brain Barrier

Journal: Frontiers in Neuroscience

doi: 10.3389/fnins.2020.00461

Effect of Mfsd2a overexpression on BBB tight junctions and vesicular transcytosis in the rat hippocampal CA1 region. (A–E) The expression of BBB permeability-related proteins, including Mfsd2a, ZO-1, claudin-5, and occludin in each group. * P
Figure Legend Snippet: Effect of Mfsd2a overexpression on BBB tight junctions and vesicular transcytosis in the rat hippocampal CA1 region. (A–E) The expression of BBB permeability-related proteins, including Mfsd2a, ZO-1, claudin-5, and occludin in each group. * P

Techniques Used: Over Expression, Expressing, Permeability

38) Product Images from "Pomegranate prevents binge alcohol-induced gut leakiness and hepatic inflammation by suppressing oxidative and nitrative stress"

Article Title: Pomegranate prevents binge alcohol-induced gut leakiness and hepatic inflammation by suppressing oxidative and nitrative stress

Journal: Redox Biology

doi: 10.1016/j.redox.2018.07.012

POM prevented the changes in gut TJ/AJ proteins and apoptosis marker proteins in binge alcohol-exposed rats. Representative levels of (A) gut TJ proteins ZO-1, claudin-1, claudin-3, and occludin, (B) AJ proteins (β-catenin and E-cadherin), plakoglobin and α-tubulin, (C) nitrated or ubiquitin-conjugated proteins, (D) apoptosis marker proteins p-JNK, Bax, and cleaved caspase-3, and (E) TUNEL analyses in the indicated groups are shown. The arrows indicate TUNEL positive cells. Densitometric quantitation of the immunoblots for each protein relative to β-actin is shown. (C) The same amounts of intestinal proteins equally combined from 4 rats/group were immunoprecipitated with the specific antibody to claudin-1 and then subjected to immunoblot analysis with the specific antibody to 3-NT (top panel) and ubiquitin (bottom). Significant difference between various treatments was determined by one-way ANOVA. Labeled characters without a common letter represent significant differences from the other group(s).
Figure Legend Snippet: POM prevented the changes in gut TJ/AJ proteins and apoptosis marker proteins in binge alcohol-exposed rats. Representative levels of (A) gut TJ proteins ZO-1, claudin-1, claudin-3, and occludin, (B) AJ proteins (β-catenin and E-cadherin), plakoglobin and α-tubulin, (C) nitrated or ubiquitin-conjugated proteins, (D) apoptosis marker proteins p-JNK, Bax, and cleaved caspase-3, and (E) TUNEL analyses in the indicated groups are shown. The arrows indicate TUNEL positive cells. Densitometric quantitation of the immunoblots for each protein relative to β-actin is shown. (C) The same amounts of intestinal proteins equally combined from 4 rats/group were immunoprecipitated with the specific antibody to claudin-1 and then subjected to immunoblot analysis with the specific antibody to 3-NT (top panel) and ubiquitin (bottom). Significant difference between various treatments was determined by one-way ANOVA. Labeled characters without a common letter represent significant differences from the other group(s).

Techniques Used: Marker, TUNEL Assay, Quantitation Assay, Western Blot, Immunoprecipitation, Labeling

39) Product Images from "Acute and Chronic Deficits in the Urinary Bladder after Spinal Contusion Injury in the Adult Rat"

Article Title: Acute and Chronic Deficits in the Urinary Bladder after Spinal Contusion Injury in the Adult Rat

Journal: Journal of Neurotrauma

doi: 10.1089/neu.2009.0997

Western blot analysis of junctional proteins in bladder tissue from injured and sham animals. ( A ) Composite images of representative Western blots of zonula occludens-1 (ZO-1) (220 kDa), occludin (65 kDa), claudin-5 (22 kDa), and β-actin (42 kDa), which were identified by binding to specific antibodies. β-Actin was used as an internal control for equal loading. ( B ) ZO-1 expression levels were significantly decreased at 2 ( p
Figure Legend Snippet: Western blot analysis of junctional proteins in bladder tissue from injured and sham animals. ( A ) Composite images of representative Western blots of zonula occludens-1 (ZO-1) (220 kDa), occludin (65 kDa), claudin-5 (22 kDa), and β-actin (42 kDa), which were identified by binding to specific antibodies. β-Actin was used as an internal control for equal loading. ( B ) ZO-1 expression levels were significantly decreased at 2 ( p

Techniques Used: Western Blot, Binding Assay, Expressing

40) Product Images from "Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway"

Article Title: Total polysaccharides of the Sijunzi decoction attenuate tumor necrosis factor-α-induced damage to the barrier function of a Caco-2 cell monolayer via the nuclear factor-κB-myosin light chain kinase-myosin light chain pathway

Journal: World Journal of Gastroenterology

doi: 10.3748/wjg.v24.i26.2867

Total polysaccharides of the Sijunzi decoction modified the expression of claudin 1, claudin 2, zo3, and occludin. A: A representative western blot of claudin 1 expression in TNF-α-damaged Caco2 cells treated with TPSJ for 12 h, 24 h, and 48 h; B: Quantification of the amounts of claudin 2 relative to GAPDH at different time points; C: A representative western blot of claudin 2 in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h; D: Quantification of the amounts of claudin 2 relative to GAPDH at different time points; E: A representative western blot of zo3 in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h; F: Quantification of the amounts of zo3 relative to GAPDH at different time points; G: A representative western blot of occludin in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h. H: Quantification of the amounts of occludin relative to GAPDH at different time points. Data are shown as the means ± standard errors of the means of at least three independent experiments ( a P
Figure Legend Snippet: Total polysaccharides of the Sijunzi decoction modified the expression of claudin 1, claudin 2, zo3, and occludin. A: A representative western blot of claudin 1 expression in TNF-α-damaged Caco2 cells treated with TPSJ for 12 h, 24 h, and 48 h; B: Quantification of the amounts of claudin 2 relative to GAPDH at different time points; C: A representative western blot of claudin 2 in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h; D: Quantification of the amounts of claudin 2 relative to GAPDH at different time points; E: A representative western blot of zo3 in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h; F: Quantification of the amounts of zo3 relative to GAPDH at different time points; G: A representative western blot of occludin in TNF-α-damaged Caco-2 cells treated with TPSJ for 12 h, 24 h, and 48 h. H: Quantification of the amounts of occludin relative to GAPDH at different time points. Data are shown as the means ± standard errors of the means of at least three independent experiments ( a P

Techniques Used: Modification, Expressing, Western Blot

Immunofluorescence analysis of the effects of total polysaccharides of the Sijunzi decoction on the tight junction proteins claudin 1, claudin 2, zo3, and occludin in tumor necrosis factor-α-damaged Caco-2 cells. Results are representative of three independent experiments. Original magnification = 400 ×.
Figure Legend Snippet: Immunofluorescence analysis of the effects of total polysaccharides of the Sijunzi decoction on the tight junction proteins claudin 1, claudin 2, zo3, and occludin in tumor necrosis factor-α-damaged Caco-2 cells. Results are representative of three independent experiments. Original magnification = 400 ×.

Techniques Used: Immunofluorescence

Related Articles

Modification:

Article Title: Establishment of an immortalized intestinal epithelial cell line from tree shrews by lentivirus-mediated hTERT gene transduction
Article Snippet: .. The following reagents were used: dithiothreitol (DTT) (Biosharp), collagenase XI (catalog # C7657, Sigma-Aldrich), insulin-transferrin-selenium (ITS-G; Life), epidermal growth factor (EGF; PeproTech), neutral protease I (Solarbio), D-sorbitol (Biosharp), pHBLV-CMVIE-ZsGreen-Puro vector (Shenggong Engineering Co., Ltd.), Escherichia coli DH5α (Invitrogen), trypsin (Thermo), Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo), phosphate buffered saline (PBS; HyClone), TaKaRa One Step PrimeScript RT-PCR Kit (Perfect Real Time) (Dalian Bao Bioengineering Co., Ltd.), RNeasy Mini Kit (Qiagen), PrimeScript II 1st Strand cDNA Synthesis Kit (Dalian Bao Bioengineering Co., Ltd.), Golden Green Mix (TsingKe), rabbit anti-cytokeratin 18 antibody (Abcam), anti-occludin antibody (Abcam), anti-β-actin antibody (Abcam), and puromycin (Solarbio). .. Isolation and culture of tree shrew intestinal epithelial cells Two-day-old tree shrews were euthanized by injecting an overdose of pentobarbital sodium.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Establishment of an immortalized intestinal epithelial cell line from tree shrews by lentivirus-mediated hTERT gene transduction
Article Snippet: .. The following reagents were used: dithiothreitol (DTT) (Biosharp), collagenase XI (catalog # C7657, Sigma-Aldrich), insulin-transferrin-selenium (ITS-G; Life), epidermal growth factor (EGF; PeproTech), neutral protease I (Solarbio), D-sorbitol (Biosharp), pHBLV-CMVIE-ZsGreen-Puro vector (Shenggong Engineering Co., Ltd.), Escherichia coli DH5α (Invitrogen), trypsin (Thermo), Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo), phosphate buffered saline (PBS; HyClone), TaKaRa One Step PrimeScript RT-PCR Kit (Perfect Real Time) (Dalian Bao Bioengineering Co., Ltd.), RNeasy Mini Kit (Qiagen), PrimeScript II 1st Strand cDNA Synthesis Kit (Dalian Bao Bioengineering Co., Ltd.), Golden Green Mix (TsingKe), rabbit anti-cytokeratin 18 antibody (Abcam), anti-occludin antibody (Abcam), anti-β-actin antibody (Abcam), and puromycin (Solarbio). .. Isolation and culture of tree shrew intestinal epithelial cells Two-day-old tree shrews were euthanized by injecting an overdose of pentobarbital sodium.

Incubation:

Article Title: Effects of imipenem combined with low-dose cyclophosphamide on the intestinal barrier in septic rats
Article Snippet: .. Membranes were subsequently incubated with primary antibodies occludin (1:5,000; cat. no. ab167161), claudin-2 (1:500; cat. no. ab53032; both Abcam, Cambridge, UK), ZO-1 (1:500; cat. no. sc-33725; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and β-actin (1:100; cat. no. ab8226) at 4°C overnight, followed by horseradish peroxidase-linked anti-rabbit IgG (1:10,000; cat. no. ab6734; both Abcam) secondary antibody for 2 h at room temperature. .. The films were then stored in a dark room and a chemiluminescent peroxidase substrate (cat. no. 34094; Thermo Fisher Scientific, Inc.) was applied according to the manufacturer's instructions.

Article Title: Replacement of grains with soybean hulls ameliorates SARA-induced impairment of the colonic epithelium barrier function of goats
Article Snippet: .. The membranes were incubated in the appropriate primary antibodies: Jun N-terminal kinase (JNK) (No. 9252S; Cell Signaling Technology, Danvers, MA), p-JNK (No. 9255S; Cell Signaling Technology), p38 (No. 8690S; Cell Signaling Technology), p-p38 (No. 4511S; Cell Signaling Technology), Erk1/2 (No. 4695S; Cell Signaling Technology), p-Erk1/2 (No. 4370S; Cell Signaling Technology), PKC- (No. 2056S; Cell Signaling Technology), occludin (No. ab167161; Abcam, Cambridge, United Kingdom), claudin-1 (No. ab15098; Abcam), and ZO-1 (No. ab214228; Abcam), then washed and incubated in corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies. .. Finally, the membranes were washed and visualized using an enhanced chemiluminescence (ECL) Kit (Pierce, Rockford, IL).

Article Title: Fluoxetine attenuates neuroinflammation in early brain injury after subarachnoid hemorrhage: a possible role for the regulation of TLR4/MyD88/NF-κB signaling pathway
Article Snippet: .. Then, we blocked the membranes with a nonfat dry milk buffer for 2 h, followed by incubation overnight with the following primary antibodies: polyclonal rabbit anti-NF-κB p65 (1:1000, ab16502, Abcam), monoclonal mouse anti-TLR4 (1:200, sc-293,072, Santa Cruz Biotechnology), polyclonal rabbit anti-MyD88 (1:1000, ab2064, Abcam), monoclonal rabbit anti-MMP9 (1:5000, ab137867, Abcam), polyclonal rabbit anti-ZO-1 (1:2000, ab96587, Abcam); polyclonal goat anti-claudin-5 (1:500, sc-17,668, Santa Cruz Biotechnology), monoclonal rabbit anti-occludin (1:50000, ab167161, Abcam). .. The membranes were then incubated with horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature.

Article Title: Qihuang decoction promotes the recovery of intestinal immune barrier dysfunction after gastrectomy in rats
Article Snippet: .. It was incubated at 4°C overnight after added anti-RhoA (abcam, ab54835, 1:1000); anti-Rac1 (abcam, ab33186, 1:1000); anti-Cdc42 (abcam, ab187643, 1:1000); anti-Claudin-1 (abcam, ab15098, 1:1000); anti-Claudin-5 (abcam, ab131259, 1:1000); anti-Occludin (abcam, ab167161, 1:1000); anti-ZO-1 (Thermo Fisher, RF232767, 1:500); anti-ZO-2 (abcam, ab191133, 1:1000); Anti-Claudin 5 (phospho Y217, abcam, ab172968, 1:1000); anti-Occludin (phospho S490, Zymed, 1:250); Anti-ZO-2 (Phospho T1118, Assay Biotechnology, P12-1069, 1:1000) and β-actin (1:1000) monoclonal antibodies (1:500). ..

Plasmid Preparation:

Article Title: Establishment of an immortalized intestinal epithelial cell line from tree shrews by lentivirus-mediated hTERT gene transduction
Article Snippet: .. The following reagents were used: dithiothreitol (DTT) (Biosharp), collagenase XI (catalog # C7657, Sigma-Aldrich), insulin-transferrin-selenium (ITS-G; Life), epidermal growth factor (EGF; PeproTech), neutral protease I (Solarbio), D-sorbitol (Biosharp), pHBLV-CMVIE-ZsGreen-Puro vector (Shenggong Engineering Co., Ltd.), Escherichia coli DH5α (Invitrogen), trypsin (Thermo), Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo), phosphate buffered saline (PBS; HyClone), TaKaRa One Step PrimeScript RT-PCR Kit (Perfect Real Time) (Dalian Bao Bioengineering Co., Ltd.), RNeasy Mini Kit (Qiagen), PrimeScript II 1st Strand cDNA Synthesis Kit (Dalian Bao Bioengineering Co., Ltd.), Golden Green Mix (TsingKe), rabbit anti-cytokeratin 18 antibody (Abcam), anti-occludin antibody (Abcam), anti-β-actin antibody (Abcam), and puromycin (Solarbio). .. Isolation and culture of tree shrew intestinal epithelial cells Two-day-old tree shrews were euthanized by injecting an overdose of pentobarbital sodium.

Mouse Assay:

Article Title: Anesthesia and Surgery Impair Blood–Brain Barrier and Cognitive Function in Mice
Article Snippet: .. Anti-β-catenin antibody (92 kDa, Cat: #9562, 1:1,000 dilution, Cell signaling, Danvers, MA, USA), anti-phosphorylated β-catenin antibody (Ser33/37/Thr41, 92 kDa, Cat: #9561, 1:1,000 dilution, Cell signaling), anti-claudin-1 antibody (19 kDa, Cat: ab15098, 1:1,000 dilution, Abcam, Cambridge, MA, USA), anti-occludin antibody (59 kDa, Cat: ab167161, 1:1,000 dilution, Abcam), anti-ZO-1 antibody (250 kDa, Cat: PA5-28858, 1:500 dilution, Thermo Fisher Scientific, Rockford, IL, USA), and anti-VE-cadherin (115 kDa, Cat: ab33168, 1 µg/ml, Abcam), anti-E-cadherin antibody (135 kDa, Cat: #5296, 1:1,000 dilution, Cell signaling) were used to detect the level of the proteins in the cortex and hippocampus of the 18-month-old mice. ..

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Abcam anti occludin
    LC3B was required for <t>occludin</t> internalization-induced PEDV entry in IPEC-J2 cells exposed to DON. (A) Generation of LC3B-knockout IPEC-J2 cells. (B, C) Effects of autophagy deficiency on the occludin expression and PEDV entry. IPEC-J2 cell monolayers were infected with 2 MOI PEDV and further cultured as indicated. Cell lysates were subjected to immunoblotting (B) with antibodies to occludin, PEDV-N protein, LC3B or β-actin (loading control). Cell lysates were subjected to IFA (C) with antibody to occludin (red) and plasmid to LC3B (green, white arrowheads). Cell nuclei were stained with DAPI (blue). The scale bar indicates 20 μm. The data are expressed as mean ± SD (n=3). # P
    Anti Occludin, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti occludin/product/Abcam
    Average 92 stars, based on 46 article reviews
    Price from $9.99 to $1999.99
    anti occludin - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    94
    Abcam occludin
    Confirmation of ZO-1, <t>occludin</t> and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin
    Occludin, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/occludin/product/Abcam
    Average 94 stars, based on 153 article reviews
    Price from $9.99 to $1999.99
    occludin - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    92
    Abcam rabbit anti occludin
    Adipose tissue extracts enable the EMT of CCA cells. Human cholangiocarcinoma (CCA) cell line RBE was incubated with complete growth medium or medium containing human adipose tissue extracts for 24 h. The cell morphology was observed under the invert microscopy (40×) ( a ). Protein expression of MMP2, MMP9, TGF-β1, claudin 1, <t>occludin,</t> E-cadherin, β-catenin, SNAIL, and Smad3 were analyzed by immunoblot analysis in response to adipose extracts incubation, and the band densitometry analysis was carried out relative to the loading control β-actin or GAPDH ( b , c ). To determine fatty acid metabolism, the culture medium with different adipose components prior to or after incubation with RBE cells for 24 h were collected for extracellular glycerol and nonestesterified fatty acid (NEFA) detection using colorimetric assays, with the data presented as the absorbance at 490 nm( d , e ). The intracellular lipids in adipose cultured RBE cells were visualized by the fluorescence dye Bodipy (20 μg/mL) using confocal microscopy analysis, using Hochest33342 (0.5 μg/mL) for nuclear staining ( f ). All the samples were prepared in triplicate, and all experiments were repeated at least three times. * P
    Rabbit Anti Occludin, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti occludin/product/Abcam
    Average 92 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    rabbit anti occludin - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    Image Search Results


    LC3B was required for occludin internalization-induced PEDV entry in IPEC-J2 cells exposed to DON. (A) Generation of LC3B-knockout IPEC-J2 cells. (B, C) Effects of autophagy deficiency on the occludin expression and PEDV entry. IPEC-J2 cell monolayers were infected with 2 MOI PEDV and further cultured as indicated. Cell lysates were subjected to immunoblotting (B) with antibodies to occludin, PEDV-N protein, LC3B or β-actin (loading control). Cell lysates were subjected to IFA (C) with antibody to occludin (red) and plasmid to LC3B (green, white arrowheads). Cell nuclei were stained with DAPI (blue). The scale bar indicates 20 μm. The data are expressed as mean ± SD (n=3). # P

    Journal: bioRxiv

    Article Title: Deoxynivalenol Promotes Porcine Epidemic Diarrhea Virus Infection and Aggravates Gut Barrier Injury

    doi: 10.1101/852608

    Figure Lengend Snippet: LC3B was required for occludin internalization-induced PEDV entry in IPEC-J2 cells exposed to DON. (A) Generation of LC3B-knockout IPEC-J2 cells. (B, C) Effects of autophagy deficiency on the occludin expression and PEDV entry. IPEC-J2 cell monolayers were infected with 2 MOI PEDV and further cultured as indicated. Cell lysates were subjected to immunoblotting (B) with antibodies to occludin, PEDV-N protein, LC3B or β-actin (loading control). Cell lysates were subjected to IFA (C) with antibody to occludin (red) and plasmid to LC3B (green, white arrowheads). Cell nuclei were stained with DAPI (blue). The scale bar indicates 20 μm. The data are expressed as mean ± SD (n=3). # P

    Article Snippet: Rabbit anti-claudin1, anti-occludin, anti-ZO-1 and anti-p-MTORC1/MTORC1 antibodies were purchased from Abcam (Cambridge, UK).

    Techniques: Knock-Out, Expressing, Infection, Cell Culture, Immunofluorescence, Plasmid Preparation, Staining

    Occludin internalization was required for DON-promoted PEDV entry in IPEC-J2 cells. (A, B) Effects of various concentrations DON on the expression and distribution of tight junction proteins in PEDV-infected IPEC-J2 cells. (C, D) Effects of occludin konckdown on PEDV entry in IPEC-J2 cells exposed to 0.5 μM DON. IPEC-J2 cell monolayers were infected with 2 MOI PEDV and further cultured as indicated. Cell lysates were subjected to immunoblotting (A, C) with antibodies to ZO-1, occludin, claudin-1, PEDV-N protein or β-actin (loading control) and subjected to IFA (B, D) with antibodies to occludin (green), claudin-1 (green) and PEDV-N protein (red). Cell nuclei were stained with DAPI (blue). The scale bar indicates 20 μm. The data are expressed as mean ± SD (n=3). # P

    Journal: bioRxiv

    Article Title: Deoxynivalenol Promotes Porcine Epidemic Diarrhea Virus Infection and Aggravates Gut Barrier Injury

    doi: 10.1101/852608

    Figure Lengend Snippet: Occludin internalization was required for DON-promoted PEDV entry in IPEC-J2 cells. (A, B) Effects of various concentrations DON on the expression and distribution of tight junction proteins in PEDV-infected IPEC-J2 cells. (C, D) Effects of occludin konckdown on PEDV entry in IPEC-J2 cells exposed to 0.5 μM DON. IPEC-J2 cell monolayers were infected with 2 MOI PEDV and further cultured as indicated. Cell lysates were subjected to immunoblotting (A, C) with antibodies to ZO-1, occludin, claudin-1, PEDV-N protein or β-actin (loading control) and subjected to IFA (B, D) with antibodies to occludin (green), claudin-1 (green) and PEDV-N protein (red). Cell nuclei were stained with DAPI (blue). The scale bar indicates 20 μm. The data are expressed as mean ± SD (n=3). # P

    Article Snippet: Rabbit anti-claudin1, anti-occludin, anti-ZO-1 and anti-p-MTORC1/MTORC1 antibodies were purchased from Abcam (Cambridge, UK).

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

    Schematic depicting role of LC3B during PEDV infection. DON exposure activates p38 signaling and triggers a complete autophagy in PEDV-infected IPEC-J2 cells. Approximately 2 hrs post-infection, LC3B induces occludin internalization to promote PEDV entry through its role as a positive regulator of autophagy. Later in infection (24 hpi.), the up-regulation of LC3B by DON contributes PEDV to escape innate immune via inhibiting the STING signaling phosphorylation, leading to production of large amounts of virus.

    Journal: bioRxiv

    Article Title: Deoxynivalenol Promotes Porcine Epidemic Diarrhea Virus Infection and Aggravates Gut Barrier Injury

    doi: 10.1101/852608

    Figure Lengend Snippet: Schematic depicting role of LC3B during PEDV infection. DON exposure activates p38 signaling and triggers a complete autophagy in PEDV-infected IPEC-J2 cells. Approximately 2 hrs post-infection, LC3B induces occludin internalization to promote PEDV entry through its role as a positive regulator of autophagy. Later in infection (24 hpi.), the up-regulation of LC3B by DON contributes PEDV to escape innate immune via inhibiting the STING signaling phosphorylation, leading to production of large amounts of virus.

    Article Snippet: Rabbit anti-claudin1, anti-occludin, anti-ZO-1 and anti-p-MTORC1/MTORC1 antibodies were purchased from Abcam (Cambridge, UK).

    Techniques: Infection

    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

    Confirmation of ZO-1, occludin and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin

    Journal: Molecular Medicine

    Article Title: Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

    doi: 10.2119/molmed.2011.00107

    Figure Lengend Snippet: Confirmation of ZO-1, occludin and claudin-1 downregulation in colonic tissues of patients with STR-IO compared with the “normal” colonic tissues of patients with colon cancer. (A–C) QRT-PCR showing mRNA levels of ZO-1 (A), occludin

    Article Snippet: The extracted proteins were separated by sodium dodecylsulfate– polyacrylamide gel electrophoresis and immunoblotted using primary antibody against ZO-1 (Abcam), occludin (Abcam), claudin-1 (Santa Cruz), HDC (Santa Cruz) and CP (Abcam).

    Techniques: Quantitative RT-PCR

    Decreased Expression of ZO-1, Occludin and Claudin-1 in STR-IO As Confirmed by QRT-PCR, Western Blotting and IHC

    Journal: Molecular Medicine

    Article Title: Histidine Decarboxylase Is Identified as a Potential Biomarker of Intestinal Mucosal Injury in Patients with Acute Intestinal Obstruction

    doi: 10.2119/molmed.2011.00107

    Figure Lengend Snippet: Decreased Expression of ZO-1, Occludin and Claudin-1 in STR-IO As Confirmed by QRT-PCR, Western Blotting and IHC

    Article Snippet: The extracted proteins were separated by sodium dodecylsulfate– polyacrylamide gel electrophoresis and immunoblotted using primary antibody against ZO-1 (Abcam), occludin (Abcam), claudin-1 (Santa Cruz), HDC (Santa Cruz) and CP (Abcam).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

    Adipose tissue extracts enable the EMT of CCA cells. Human cholangiocarcinoma (CCA) cell line RBE was incubated with complete growth medium or medium containing human adipose tissue extracts for 24 h. The cell morphology was observed under the invert microscopy (40×) ( a ). Protein expression of MMP2, MMP9, TGF-β1, claudin 1, occludin, E-cadherin, β-catenin, SNAIL, and Smad3 were analyzed by immunoblot analysis in response to adipose extracts incubation, and the band densitometry analysis was carried out relative to the loading control β-actin or GAPDH ( b , c ). To determine fatty acid metabolism, the culture medium with different adipose components prior to or after incubation with RBE cells for 24 h were collected for extracellular glycerol and nonestesterified fatty acid (NEFA) detection using colorimetric assays, with the data presented as the absorbance at 490 nm( d , e ). The intracellular lipids in adipose cultured RBE cells were visualized by the fluorescence dye Bodipy (20 μg/mL) using confocal microscopy analysis, using Hochest33342 (0.5 μg/mL) for nuclear staining ( f ). All the samples were prepared in triplicate, and all experiments were repeated at least three times. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Adipocytes promote cholangiocarcinoma metastasis through fatty acid binding protein 4

    doi: 10.1186/s13046-017-0641-y

    Figure Lengend Snippet: Adipose tissue extracts enable the EMT of CCA cells. Human cholangiocarcinoma (CCA) cell line RBE was incubated with complete growth medium or medium containing human adipose tissue extracts for 24 h. The cell morphology was observed under the invert microscopy (40×) ( a ). Protein expression of MMP2, MMP9, TGF-β1, claudin 1, occludin, E-cadherin, β-catenin, SNAIL, and Smad3 were analyzed by immunoblot analysis in response to adipose extracts incubation, and the band densitometry analysis was carried out relative to the loading control β-actin or GAPDH ( b , c ). To determine fatty acid metabolism, the culture medium with different adipose components prior to or after incubation with RBE cells for 24 h were collected for extracellular glycerol and nonestesterified fatty acid (NEFA) detection using colorimetric assays, with the data presented as the absorbance at 490 nm( d , e ). The intracellular lipids in adipose cultured RBE cells were visualized by the fluorescence dye Bodipy (20 μg/mL) using confocal microscopy analysis, using Hochest33342 (0.5 μg/mL) for nuclear staining ( f ). All the samples were prepared in triplicate, and all experiments were repeated at least three times. * P

    Article Snippet: The primary antibodies used were rabbit anti-FABP4 (EPR3579, Abcam, Cambridge, MA, USA, 1:1000), rabbit anti-claudin 1 (Abcam, Cambridge, MA, USA, 1:1000), rabbit anti-occludin (EPR8208, Abcam, Cambridge, MA, USA, 1:50,000), rabbit anti-E-cadherin (EP700Y, Abcam, Cambridge, MA, USA, 1:10,000), rabbit anti-SNAIL (Abcam, Cambridge, MA, USA, 1:1000), rabbit anti-Smad3 (EP568Y, Abcam, Cambridge, MA, USA, 1:1000), rabbit anti-β-catenin (Cell Signaling Technology, Danvers, MA, USA, 1:1000), rabbit anti-MMP2 (Cell Signaling Technology, Danvers, MA, USA,1:1000), rabbit anti-MMP9 (Cell Signaling Technology, Danvers, MA, USA,1:1000), and anti-β-actin (Beyotime Biotechnology, Haimen, China).

    Techniques: Incubation, Microscopy, Expressing, Cell Culture, Fluorescence, Confocal Microscopy, Staining