occludin  (Bioss)

Journal: Journal of Virology

Article Title: HIV transgenic mouse monocytes display increased in vivo migration across the blood-brain barrier associated with increased expression of genes associated with mononuclear leukocyte movement

doi: 10.1128/jvi.02063-25

Figure Lengend Snippet: Increased transmigration of HIV transgenic monocytes into the brains of recipient mice. ( A ) Evaluation of BBB integrity by immunofluorescent visualization of the ETJ proteins claudin-5, occludin, and ZO-1 in cerebral arteries 4 h after B6 mice were injected intraperitoneally with either PBS or LPS (3 mg/kg). Coronal brain sections (15-µm thick) were stained for cell nuclei (blue), claudin-5 (red), occludin (green), and ZO-1 (yellow), and were imaged at 20× magnification (scale bar = 100 µm). ( B ) Continuity of ETJ protein expression was determined by measuring the average length of blood vessels continuously stained for ETJ proteins in four different fields in similar hypothalamic regions of the mouse brain tissue for PBS and LPS-treated mice. Each symbol in the bar graph represents an average continuous ETJ expression in one visual field, with mean ± SEM shown, and the data are representative of two experiments, each done with n = 2 mice. Statistical analysis was performed using unpaired t -tests between groups. ( C ) Experimental set-up and timeline. Monocytes, harvested from JRCCC bone marrow leukocytes (BML) and highly purified by immunomagnetic sorting, were injected intravenously (20 × 10 6 monocytes/mouse) into the tail veins of B6 mice 4 h after they were intraperitoneally injected with LPS (3 mg/kg, n = 4 mice) or PBS ( n = 6 mice). ( D ) DNA from harvested mouse brains was evaluated for the number of human CD4 copies by droplet digital PCR (ddPCR), which represents the presence of individual JRCCC monocytes. Each symbol represents the number of human CD4 copies detected by ddPCR in the brain of an individual mouse. The individual and average number of human CD4 DNA in copies/brain for PBS or LPS-treated mice are shown with mean ± SEM (* P ≤ 0.05). ( E ) Two days after the highly purified JRCCC monocytes were injected intravenously into B6 mice, the mice were bled to quantify the level of HIV viremia by RT-qPCR. The data represent pooled data from five experiments, with each symbol representing the number of HIV RNA copies/mL in individual mice. The individual and average viral loads with mean ± SEM of HIV-Gag RNA copies/mL in the plasma of PBS or LPS-treated mice are shown.

Article Snippet: Tissue sections were then blocked in 0.1 M PBS containing 10% horse serum and 3% Triton X-100 for 1 h at room temperature, followed by overnight incubation at 4°C with the following primary antibodies, claudin-5 polyclonal rabbit antibody (1:550, Thermo Fisher, #34-1600), ZO-1 monoclonal rat antibody (1:300, Thermo Fisher, #14-9776-82), or occludin polyclonal guinea pig antibody (1:50, Origene, #AP26410PU-N).

Techniques: Transmigration Assay, Transgenic Assay, Injection, Staining, Expressing, Purification, Digital PCR, Quantitative RT-PCR, Clinical Proteomics

Journal: Animal Nutrition

Article Title: Engineered Bacillus subtilis WB600/ZD reduces post-weaning diarrhea in piglets by modulating gut microbiota and aryl hydrocarbon receptor (AHR) signaling

doi: 10.1016/j.aninu.2025.09.008

Figure Lengend Snippet: WB600/ZD-FMT and AHR activation alleviate ileal barrier damage caused by Salmonella enterica subsp . enterica serovar Infantis( S. Infantis) infection. (A) Immunohistochemical detection of mucins2 (MUC2) expression in the ileum (the black arrow). (B) Immunofluorescence detection of zonula occludens-1 (ZO-1) expression in the ileum. (C) Claudin-1 and occludin expression in the ileum by Western blot assays. (D-F) RT-qPCR detection of the expression of MUC2 and mucin-related genes trefoil factor 3 ( TFF3 ) and Galactose-3-O-sulfotransferase 2 ( GAL3ST2 ) in the ileum. All data are presented as means ± SD from at least three independent experiments. The P -values shown denote statistical significance for all pairwise comparisons between groups. WB600/ZD is an engineered Bacillus subtilis strain (derived from the protease-deficient parental strain WB600) expressing the antimicrobial defensin peptide from Zophobas atratus , where “ZD” denotes the Zophobas atratus defensin. FMT = fecal microbiota transplantation; SI = S . Infantis infection; CH CH223191. ABX = antibiotic mixture; FICZ = 6-formylindolo[3,2-b]carbazole; PWD = post-weaning diarrhea; AHR = aryl hydrocarbon receptor; CYP1A1 = cytochrome P450 family 1 subfamily A member 1.

Article Snippet: The following antibodies were used for Western blot: AHR polyclonal antibody (AF6278, 1:500; Affinity Biosciences Ltd., Nanjing, China), CYP1A1 polyclonal antibody (AF5312, 1:500; Affinity Biosciences Ltd.), β-actin monoclonal antibody (66009-1-Ig, 1:8000; Proteintech Group, Inc., Wuhan, Hubei, China), claudin-1 polyclonal antibody (28674-1-AP, 1:5000; Proteintech Group, Inc.), occludin polyclonal antibody (27260-1-AP, 1:5000; Proteintech Group, Inc.).

Techniques: Activation Assay, Infection, Immunohistochemical staining, Expressing, Immunofluorescence, Western Blot, Quantitative RT-PCR, Derivative Assay, Transplantation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Superior thermotolerance in young versus adult rats undergoing heat stroke is associated with age-related differences in intestinal barrier integrity and heat shock protein responses

doi: 10.3389/fcell.2026.1642359

Figure Lengend Snippet: Western blot and immunofluorescence analyses reveal that young rats exhibit less disruption in the expression and distribution of intestinal barrier proteins compared to adult rats following HS. (A) Representative western blots showing ZO-1, E-cadherin, and Occludin expression after HS. GAPDH was used as the loading control. (B–D) Quantification of band intensities for ZO-1 (B) , E-cadherin (C) , and Occludin (D) , normalized to GAPDH. (E) Cell nuclei were stained with DAPI (blue), and ZO-1 (red), E-cadherin (green), and Occludin (yellow) were visualized using specific antibodies (scale bar, 100 μm). Data are presented as mean ± SEM, n = 3 per group. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Membranes were blocked with quick blocking buffer (Beyotime Institute of Biotechnology, P0252, Shanghai, China) for 1 h at room temperature, followed by overnight incubation at 4 °C with the following primary antibodies: ZO-1 (Proteintech, 21773-1-AP, rabbit, 1:5,000), E-cadherin (Proteintech, 20874-1-AP, rabbit, 1:20,000), Occludin (Proteintech, 27260-1-AP, rabbit, 1:5,000), HSP90 (Proteintech, 13171-1-AP, rabbit, 1:6,000), HSP70 (Proteintech, 10995-1-AP, rabbit, 1:10,000), HSP60 (Proteintech, 15282-1-AP, rabbit, 1:6,000), HSP40 (Proteintech, 13174-1-AP, rabbit, 1:10,000), and GAPDH (Proteintech, 60004-1-Ig, mouse, 1:50,000).

Techniques: Western Blot, Immunofluorescence, Disruption, Expressing, Control, Staining