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mouse anti human icam (R&D Systems)

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R&D Systems mouse anti human icam
Mouse Anti Human Icam, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+anti+icam+1/Human+ICAM-1%2FCD54+Antibody/pm41944190-237-44-47
Average 93 stars, based on 110 article reviews

mouse anti human icam - by Bioz Stars, 2026-09

93/100 stars

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other:

Article Title: Chemokine CCL2 and its receptor CCR2 are increased in the hippocampus following pilocarpine-induced status epilepticus
Article Snippet: In order to decipher the cell types that express CCR2, double-labeling immunohistochemistry was performed using the following antibodies combined with CCR2 staining: neuronal nuclei protein (mouse anti-NeuN, 1:1000; Chemicon) that stains neurons; microtubule-associated protein 2 (mouse anti-MAP-2, 1:1000; Millipore), a neuron-specific microtubule protein; glial fibrillary acidic protein (mouse anti-GFAP-Cy3, 1:500; Sigma), a marker of astrocytes; ionized calcium binding adaptor molecule 1 (goat anti-Iba1, 1:500; Abcam) that stains macrophage/microglia and a marker of endothelial cells (mouse anti-ICAM-1, 1:200, R&D Systems).

Western Blot:

Article Title: Endothelial cells elicit a pro-inflammatory response to SARS-CoV-2 without productive viral infection
Article Snippet: .. The following antibodies were used: mouse anti-ACE2 (Santa Cruz, sc-390851, IF1:200) goat anti-ACE2 (R&D systems, AF933, WB 1:1000), mouse anti-dsRNA (Millipore, MABE1134, IF1:100), rabbit anti-SARS-CoV nucleocapsid protein/NP (Sino Biological, 40143-R040, WB1:1000 IF1:200), rabbit anti-TMPRSS2 (Abcam, ab92323, WB1:1000), rabbit anti-cleaved caspase 3 (Cell Signaling, 9661 IF1:30 after pre-labeling), rabbit anti-GAPDH (Cell Signaling, 2118, WB1:2000), mouse anti-ICAM-1 (R&D systems, BBA3, IF1:200), mouse anti-ICAM-1 (Santa Cruz Biotechnology, sc-8439, WB1:1000), Phalloidin-Alexa555 (Cytoskeleton, PHDH1-A, IF1:500), Phalloidin-Alexa670 (Cytoskeleton, PHDN1-A, IF1:200). ..

Incubation:

Article Title: Montelukast reduces grey matter abnormalities and functional deficits in a mouse model of inflammation-induced encephalopathy of prematurity
Article Snippet: .. Sections were incubated overnight (4 °C) in one of the following primary antibodies, diluted in 1% NHS block: Goat anti-serum albumin (1:5000, Abcam), rabbit anti-parvalbumin (PV, 1:200, Abcam), mouse anti-MBP (1:200, Millipore), mouse-anti-CNPase (1:200, Neomarkers), mouse anti-ICAM-1 (1:200, R&D Systems), goat anti-GFAP (1:300, Abcam) and Lycopersicon esculentum (tomato) lectin (1:200, Vector, UK). .. Biotinylated secondary antibodies (1:200, Vector, diluted in 1% NHS block, 2 h, RT) were used, followed by HRP (ABC Elite, Vector, UK) and DAB prior to imaging and analysis.

Blocking Assay:

Immunostaining:

Article Title: Modeling alpha-synuclein pathology in a human brain-chip to assess blood-brain barrier disruption
Article Snippet: .. Immunostaining was performed with specific primary antibodies (Supplementary Table ): rabbit anti-TH (1:500;abcam, ab6211), chicken anti-TH (1:100;abcam, ab76442), goat anti-GFAP (1:300;abcam, ab53554), mouse anti-TH (1:100;Sigma, T2928), Rabbit anti-FOXA2 (1:200;Cell Signaling, 8186), rabbit anti-LMX1A (1:100;Sigma, ZRB1373), mouse ant-CD68 (1:100;abcam, ab955), mouse anti-ICAM-1 (1:100;R&D Systems, NET30), rabbit MAP2 (1:100;abcam, ab32454), mouse anti-TOM20 (1:100;abcam, ab56783), rabbit anti-IBA1 (1:50;FCDI, 019-19741), mouse anti-phosphoSer129 (1:100;abcam, ab184674), rabbit anti-phosphoSer129 (1:100;Cell Signaling, 23706), rabbit anti-ki67 (1:10;abcam, ab197234), rabbit anti-CD68 (1:100;abcam, ab213363), rabbit anti-Cleaved Caspase-3 (1:100;abcam, ab32042), rabbit anti-NG2 (1:100;abcam, ab83178), rabbit anti-TMEM119 (1:100;abcam, ab185333), rabbit anti-pSer129-αSyn (1:100;abcam ab51253), mouse anti-MAP2 (1:200;Thermo Fisher Scientific, MA512823), rabbit anti-CD31 (1:25;Thermo Fisher Scientific, RB-10333-P1), mouse anti-Claudin-1 (1:25;Thermo Fisher Scientific, 37-4900), mouse anti-Claudin-5 (1:50;Thermo Fisher Scientific, 35-2500), mouse anti-Occludin (1:100;Thermo Fisher Scientific, 33-1500), rabbit anti-ZO-1 (1:200;Thermo Fisher Scientific, 40-2200), mouse anti-ZO-1 (1:200;Thermo Fisher Scientific, ZO1-1A12). ..

Purification:

Article Title: Shear forces induce ICAM-1 nanoclustering on endothelial cells that impact on T-cell migration.
Article Snippet: Secondary Ab goat-anti-mouse-AF488 (Cat#11001, Lot# 2015565), donkeyanti-mouseAlexa 647 (Cat#A32787, Lot#UI291059), goat-anti-rabbit Alexa 647 (Cat# A21244), and donkey-anti-rabbit Alexa 647 (Cat# A32795, Lot# UG289708) were from Invitrogen. .. Purified ICAM-1 Fc Chimera (Cat# 721- IC, Lot# DLA1119041, stock 50 mg/mL) and mouse-anti-ICAM-1 (BBIGI1, Cat# BBA3, Lot# ANE1319121) used for cross-linking experiments on ECs were from R&D systems (Minneapolis, MN). .. Polyclonal rabbit-antiICAM-1 (H-108, Cat# SC7891, Lot# H1712, stock 200 mg/mL) was from Santa Cruz (Dallas, TX).

Image Search Results

(A-D) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and cells were fixed at 24, 48, and 72h post-infection. A) Immunofluorescence staining of ICAM-1 (green), F-actin (phalloidin; grey), and nuclei (DAPI; blue) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. B) Quantification of HMVEC-L ICAM-1 intensity analysed by 2-way ANOVA with Sidak’s multiple comparison’s test. C) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. D) Quantification of the percentage of gaps in the endothelial monolayer was analysed by 2-way ANOVA with Sidak’s multiple comparison’s test. E-H) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and cells were fixed at 72h post-infection. E) Immunofluorescence staining of Zombie Red (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of four independent experiments. F) The percentage of Zombie Red-positive cells between conditions was analysed using the Kruskal-Wallis test with Dunn’s multiple testing correction. G) Immunofluorescence staining and imaging of CellTrace Yellow-labelled platelets (yellow), VE-cadherin (magenta), and nuclei (DAPI; blue) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. H) Gaps in the cellular monolayer were outlined relative to VE-cadherin staining, and only the platelets present in gaps were quantified. I) The number of platelets (CellTrace Yellow positive particles larger than 2 mm) in gaps was quantified and analysed by the Kruskal-Wallis test with Dunn’s multiple testing correction. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented by the large dot (colour-coded by experiment), and the data show the mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: (A-D) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and cells were fixed at 24, 48, and 72h post-infection. A) Immunofluorescence staining of ICAM-1 (green), F-actin (phalloidin; grey), and nuclei (DAPI; blue) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. B) Quantification of HMVEC-L ICAM-1 intensity analysed by 2-way ANOVA with Sidak’s multiple comparison’s test. C) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. D) Quantification of the percentage of gaps in the endothelial monolayer was analysed by 2-way ANOVA with Sidak’s multiple comparison’s test. E-H) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and cells were fixed at 72h post-infection. E) Immunofluorescence staining of Zombie Red (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of four independent experiments. F) The percentage of Zombie Red-positive cells between conditions was analysed using the Kruskal-Wallis test with Dunn’s multiple testing correction. G) Immunofluorescence staining and imaging of CellTrace Yellow-labelled platelets (yellow), VE-cadherin (magenta), and nuclei (DAPI; blue) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. H) Gaps in the cellular monolayer were outlined relative to VE-cadherin staining, and only the platelets present in gaps were quantified. I) The number of platelets (CellTrace Yellow positive particles larger than 2 mm) in gaps was quantified and analysed by the Kruskal-Wallis test with Dunn’s multiple testing correction. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented by the large dot (colour-coded by experiment), and the data show the mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against mouse ICAM-1 (BioXCell; BE0020-1) and mouse CD31 (New England Biolabs; 77699S).

Techniques: Infection, Immunofluorescence, Staining, Co-Culture Assay, Imaging

(A-E) NHBE/HMVEC-L co-cultures infected with SARS-CoV-2 (MOI 1) and then treated with 100 mg/mL dexamethasone or media alone immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L in the basal compartment of the co-culture. Images are representative of 2 independent experiments. B) Quantification of HMVEC-L ICAM-1 intensity, where data shows mean + SD. C) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. D) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. E) Viral titres from the apical compartment of SARS-CoV-2-infected NHBE/HMVEC-L co-cultures, untreated or treated with 100 mg /mL dexamethasone, at 72h post-infection. n = 3 independent experiments, analysed by unpaired, two-way t -test. F) Schematic of supernatant transfer experiment. G-J) NHBE monocultures were infected with SARS-CoV-2 for 48h. The supernatant from the basal compartment was then transferred onto HMVEC-L. 100 mg /mL dexamethasone or PBS was added to the NHBE basal supernatants before they were transferred onto HMVEC-L. After 24h, HMVEC-L were fixed for immunofluorescence staining. G) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L. Images are representative of 3 independent experiments. H) Quantification of HMVEC-L ICAM-1 intensity analysed by one-way ANOVA with Sidak’s multiple comparison’s test. I) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L. Images are representative of three independent experiments. J) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented by the large dot (colour-coded by experiment), and the data show the mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: (A-E) NHBE/HMVEC-L co-cultures infected with SARS-CoV-2 (MOI 1) and then treated with 100 mg/mL dexamethasone or media alone immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L in the basal compartment of the co-culture. Images are representative of 2 independent experiments. B) Quantification of HMVEC-L ICAM-1 intensity, where data shows mean + SD. C) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L in the basal compartment of the co-culture. Images are representative of three independent experiments. D) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. E) Viral titres from the apical compartment of SARS-CoV-2-infected NHBE/HMVEC-L co-cultures, untreated or treated with 100 mg /mL dexamethasone, at 72h post-infection. n = 3 independent experiments, analysed by unpaired, two-way t -test. F) Schematic of supernatant transfer experiment. G-J) NHBE monocultures were infected with SARS-CoV-2 for 48h. The supernatant from the basal compartment was then transferred onto HMVEC-L. 100 mg /mL dexamethasone or PBS was added to the NHBE basal supernatants before they were transferred onto HMVEC-L. After 24h, HMVEC-L were fixed for immunofluorescence staining. G) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L. Images are representative of 3 independent experiments. H) Quantification of HMVEC-L ICAM-1 intensity analysed by one-way ANOVA with Sidak’s multiple comparison’s test. I) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L. Images are representative of three independent experiments. J) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented by the large dot (colour-coded by experiment), and the data show the mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against mouse ICAM-1 (BioXCell; BE0020-1) and mouse CD31 (New England Biolabs; 77699S).

Techniques: Infection, Immunofluorescence, Staining, Co-Culture Assay

(A-H) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected and treated with 10 mg/mL anti-TNF (Adalimumab) immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining for ICAM-1 (green), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. B) ICAM-1 intensity between conditions was analysed by one-way ANOVA, with Sidak’s multiple comparison test. C) Immunofluorescence staining for VE-cadherin (magenta), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. D) Quantification of gaps in the endothelial monolayer under different conditions was determined by calculating the percentage of the image area covered by gaps, and analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. E) Zombie Red-stained cells (magenta) indicate cells (containing F-actin and nuclei) undergoing cell death. Images are representative of n = 3 independent experiments. F) The percentage of Zombie Red positive cells was analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. G) Immunofluorescent staining of CellTrace Yellow-labelled platelets incubated with HMVEC-L. Images are representative of n = 3 independent experiments. H) The number of platelets (CellTrace Yellow positive particles larger than 2 mm) in gaps was quantified and analysed by the Kruskal-Wallis test with Dunn’s multiple testing correction. I) NHBE monocultures and NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and TNF levels in the apical and basal supernatants were analysed at 24, 48, and 72h post-infection. Data show the mean ± SEM of 3 independent experiments, analysed by 2-way ANOVA. (J-M) NHBE monocultures were infected with SARS-CoV-2 for 48h. The supernatant from the basal compartment was then transferred onto HMVEC-L. Anti-TNF (10 mg/mL) or PBS was added to the NHBE basal supernatants before they were transferred onto HMVEC-L. After 24h, HMVEC-L were fixed for immunofluorescence staining. J) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L. Images are representative of 3 independent experiments. K) Quantification of HMVEC-L ICAM-1 intensity was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. L) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L. Images are representative of three independent experiments. M) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented with the large dot (colour-coded per experiment), and the data shows mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: (A-H) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected and treated with 10 mg/mL anti-TNF (Adalimumab) immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining for ICAM-1 (green), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. B) ICAM-1 intensity between conditions was analysed by one-way ANOVA, with Sidak’s multiple comparison test. C) Immunofluorescence staining for VE-cadherin (magenta), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. D) Quantification of gaps in the endothelial monolayer under different conditions was determined by calculating the percentage of the image area covered by gaps, and analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. E) Zombie Red-stained cells (magenta) indicate cells (containing F-actin and nuclei) undergoing cell death. Images are representative of n = 3 independent experiments. F) The percentage of Zombie Red positive cells was analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. G) Immunofluorescent staining of CellTrace Yellow-labelled platelets incubated with HMVEC-L. Images are representative of n = 3 independent experiments. H) The number of platelets (CellTrace Yellow positive particles larger than 2 mm) in gaps was quantified and analysed by the Kruskal-Wallis test with Dunn’s multiple testing correction. I) NHBE monocultures and NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected, and TNF levels in the apical and basal supernatants were analysed at 24, 48, and 72h post-infection. Data show the mean ± SEM of 3 independent experiments, analysed by 2-way ANOVA. (J-M) NHBE monocultures were infected with SARS-CoV-2 for 48h. The supernatant from the basal compartment was then transferred onto HMVEC-L. Anti-TNF (10 mg/mL) or PBS was added to the NHBE basal supernatants before they were transferred onto HMVEC-L. After 24h, HMVEC-L were fixed for immunofluorescence staining. J) Immunofluorescence staining of ICAM-1 (green) in HMVEC-L. Images are representative of 3 independent experiments. K) Quantification of HMVEC-L ICAM-1 intensity was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. L) Immunofluorescence staining of VE-cadherin (magenta) in HMVEC-L. Images are representative of three independent experiments. M) Quantification of the percentage of gaps in the endothelial monolayer was analysed by one-way ANOVA with Sidak’s multiple comparison’s test. Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented with the large dot (colour-coded per experiment), and the data shows mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against mouse ICAM-1 (BioXCell; BE0020-1) and mouse CD31 (New England Biolabs; 77699S).

Techniques: Infection, Immunofluorescence, Staining, Comparison, Incubation

(A-F) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected and treated with 10 mg/mL Anakinra immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining for ICAM-1 (green), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 5 independent experiments. B) ICAM-1 intensity between conditions was analysed by one-way ANOVA, with Sidak’s multiple comparison test. C) Immunofluorescence staining for VE-cadherin (magenta), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. D) Quantification of gaps in the endothelial monolayer under different conditions was determined by calculating the percentage of the image area covered by gaps, and analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. E) Zombie Red-stained cells (magenta) indicate cells (containing F-actin and nuclei) undergoing cell death. Images are representative of n = 3 independent experiments. F) The percentage of Zombie Red positive cells was analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. G) IL-1β levels in the apical supernatant of NHBE monocultures infected with SARS-CoV-2 (MOI 1) or mock-infected, at 24, 48, and 72h post-infection (n = 3 independent experiments). H) TNF levels in the apical (left panel) and basal (right panel) supernatant of SARS-CoV-2-infected NHBE/HMVEC-L co-cultures, untreated or treated with Anakinra, at 24, 48, and 72h post-infection (n = 2 independent experiments). Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented with the large dot (colour-coded per experiment), and the data shows mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: (A-F) NHBE/HMVEC-L co-cultures were infected with SARS-CoV-2 (MOI 1) or mock-infected and treated with 10 mg/mL Anakinra immediately post-infection. Cells were fixed at 72h post-infection. A) Immunofluorescence staining for ICAM-1 (green), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 5 independent experiments. B) ICAM-1 intensity between conditions was analysed by one-way ANOVA, with Sidak’s multiple comparison test. C) Immunofluorescence staining for VE-cadherin (magenta), F-actin (Phalloidin; grey), and nuclei (DAPI; blue). Images are representative of n = 3 independent experiments. D) Quantification of gaps in the endothelial monolayer under different conditions was determined by calculating the percentage of the image area covered by gaps, and analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. E) Zombie Red-stained cells (magenta) indicate cells (containing F-actin and nuclei) undergoing cell death. Images are representative of n = 3 independent experiments. F) The percentage of Zombie Red positive cells was analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. G) IL-1β levels in the apical supernatant of NHBE monocultures infected with SARS-CoV-2 (MOI 1) or mock-infected, at 24, 48, and 72h post-infection (n = 3 independent experiments). H) TNF levels in the apical (left panel) and basal (right panel) supernatant of SARS-CoV-2-infected NHBE/HMVEC-L co-cultures, untreated or treated with Anakinra, at 24, 48, and 72h post-infection (n = 2 independent experiments). Scale bar for all images = 50 µm. 5 ROIs per experiment were quantified (small dots) and are colour-coded per experiment. The average of the 5 ROIs is represented with the large dot (colour-coded per experiment), and the data shows mean ± SEM. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against mouse ICAM-1 (BioXCell; BE0020-1) and mouse CD31 (New England Biolabs; 77699S).

Techniques: Infection, Immunofluorescence, Staining, Comparison

A) Representative immunohistochemistry (IHC) images of lungs from SARS-CoV-2-infected (10 4 TCID50) or mock-infected wild-type (WT), Tnf -/- , Il1b -/- , and Tnf -/- IL1b -/- mice, harvested at 3 days post-infection (dpi). Tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 50 µm. Images are representative of 5 images per mouse, 3 mice per group. B) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by 2-way ANOVA with Sidak’s multiple testing correction. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented with the large dot (colour-coded per mouse), and the data shows mean ± SEM. C-F) Aged K18-hACE c57BL/6□J mice infected with SARS-CoV-2 (10 3 PFU) and treated with an isotype control or anti-IL-1β antibody at 1h or 3 days post-infection. Lungs were harvested at day 6 post-infection. C) Lung tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 100 µm. Images are representative of 5 images per mouse, 4-6 mice per group. D) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented by the large dot (colour-coded per mouse), and the data show the mean ± SEM. E) Lung tissues were stained for fibrinogen. Scale bar = 50 µm. Images are representative of 4-6 mice per group. F) Quantification of fibrinogen staining intensity analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: bioRxiv

Article Title: IL-1β and TNF drive endothelial dysfunction and coagulopathy in acute COVID-19

doi: 10.64898/2026.03.21.713333

Figure Lengend Snippet: A) Representative immunohistochemistry (IHC) images of lungs from SARS-CoV-2-infected (10 4 TCID50) or mock-infected wild-type (WT), Tnf -/- , Il1b -/- , and Tnf -/- IL1b -/- mice, harvested at 3 days post-infection (dpi). Tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 50 µm. Images are representative of 5 images per mouse, 3 mice per group. B) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by 2-way ANOVA with Sidak’s multiple testing correction. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented with the large dot (colour-coded per mouse), and the data shows mean ± SEM. C-F) Aged K18-hACE c57BL/6□J mice infected with SARS-CoV-2 (10 3 PFU) and treated with an isotype control or anti-IL-1β antibody at 1h or 3 days post-infection. Lungs were harvested at day 6 post-infection. C) Lung tissues were stained with CD31 (magenta), ICAM-1 (green), and DAPI (blue). Scale bar = 100 µm. Images are representative of 5 images per mouse, 4-6 mice per group. D) Quantification of ICAM-1 intensity in areas of CD31 staining, analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. 5 ROIs per mouse were quantified (small dots) and are colour-coded per mouse. The average of the 5 ROIs is represented by the large dot (colour-coded per mouse), and the data show the mean ± SEM. E) Lung tissues were stained for fibrinogen. Scale bar = 50 µm. Images are representative of 4-6 mice per group. F) Quantification of fibrinogen staining intensity analysed by one-way ANOVA, with Sidak’s multiple comparison’s test. Asterisks indicate statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibodies against mouse ICAM-1 (BioXCell; BE0020-1) and mouse CD31 (New England Biolabs; 77699S).

Techniques: Immunohistochemistry, Infection, Staining, Control

(A) Flow cytometry analysis showing frequencies of CD31⁺ endothelial cells expressing ICAM-1, VCAM-1, P-selectin, or E-selectin from the periphery vs. core of large (∼900 mm³) YUMMER1.7 tumors. Statistical analysis was performed using a paired two-tailed Student’s t-test. (B) Expression of CD11a (LFA-1 α-subunit; Δ geometric MFI) on the indicated tumor-infiltrating immune populations. (C, D) Representative immunofluorescence showing ICAM-1 (green) on CD31⁺ vessels (red) in small (∼100 mm³) and large (∼900 mm³) tumors at periphery and core (C) and corresponding quantification of ICAM-1⁺/CD31⁺ area (%) (D). Scale bar in C, 100 µm. (E-H) Representative images of extravasated fibrin(ogen) (red) (E) and pericyte (desmin, red) (G) relative to CD31 (green) in small and large tumors at periphery and core and corresponding quantification of fibrin/CD31 area (%) (F) and desmin/CD31 area (%) (H), respectively. Scale bar for E and G, 100 µm. Statistical analysis was performed using paired or unpaired two-tailed Student’s t-test. Each point represents one mouse; bars show ± SEM.

Journal: bioRxiv

Article Title: Spatial polarization of endothelial ICAM-1 governs T-cell exclusion in melanoma

doi: 10.64898/2026.03.19.712709

Figure Lengend Snippet: (A) Flow cytometry analysis showing frequencies of CD31⁺ endothelial cells expressing ICAM-1, VCAM-1, P-selectin, or E-selectin from the periphery vs. core of large (∼900 mm³) YUMMER1.7 tumors. Statistical analysis was performed using a paired two-tailed Student’s t-test. (B) Expression of CD11a (LFA-1 α-subunit; Δ geometric MFI) on the indicated tumor-infiltrating immune populations. (C, D) Representative immunofluorescence showing ICAM-1 (green) on CD31⁺ vessels (red) in small (∼100 mm³) and large (∼900 mm³) tumors at periphery and core (C) and corresponding quantification of ICAM-1⁺/CD31⁺ area (%) (D). Scale bar in C, 100 µm. (E-H) Representative images of extravasated fibrin(ogen) (red) (E) and pericyte (desmin, red) (G) relative to CD31 (green) in small and large tumors at periphery and core and corresponding quantification of fibrin/CD31 area (%) (F) and desmin/CD31 area (%) (H), respectively. Scale bar for E and G, 100 µm. Statistical analysis was performed using paired or unpaired two-tailed Student’s t-test. Each point represents one mouse; bars show ± SEM.

Article Snippet: Mice received intraperitoneal injections of anti-mouse ICAM-1 (CD54) monoclonal antibody (BioXcell, BE0020, clone YN1/1.7.4; 2.5 mg/kg), anti-PD-1 (BioXcell, BP0146, clone RMP1-14; 5 mg/kg), or rat IgG2b isotype control (BioXcell, BE0090) every other day for a total of seven doses.

Techniques: Flow Cytometry, Expressing, Two Tailed Test, Immunofluorescence

(A) Schematic diagram of treatment. 2.5 × 10 5 YUMMER1.7 cells were subcutaneously inoculated in C57BL/6 mice. When tumors reached ∼70 mm³ (days 8–10), mice were treated with anti-ICAM-1 or rat IgG isotype control (2.5 mg/kg, i.p.) every other day for seven doses; tumors were collected on days 21–24. (B-C) Tumor growth curves showing group means ± SEM (B) and individual trajectories (C) for IgG (black, n = 10) and anti-ICAM-1 (blue, n = 10) pooled from three independent experiments. P -value at the final time point by two-way ANOVA. (D) Representative immunofluorescence showing CD8⁺ T-cell distribution (green) across periphery to core in IgG- and anti-ICAM-1–treated tumors. Solid line marks the tumor boundary, and dashed line indicates the periphery–core boundary. Scale bar, 200 µm. (E) Quantification of CD8⁺ T-cell counts in periphery vs core (IgG, n = 6; anti-ICAM-1, n = 6). Statistical analysis was performed using unpaired or paired student two-tailed Student’s t-test. ( F-J ) Flow cytometric analysis of whole tumors comparing the frequency of CD8 + T cells among live cells (F), Granzyme B + cells among CD8 + T cells (G), and CD45 + immune cells among live cells (H), CD4 + T cells (I) among live cells, and regulatory T cells (Treg; CD4 + Foxp3 + ) among CD4 + T cells (J) between IgG (black, n = 14) and anti-ICAM-1 (blue, n =14). Statistical analysis was performed unpaired two-tailed Student’s t-test. Each point represents one mouse; bars show mean ± SEM.

Journal: bioRxiv

Article Title: Spatial polarization of endothelial ICAM-1 governs T-cell exclusion in melanoma

doi: 10.64898/2026.03.19.712709

Figure Lengend Snippet: (A) Schematic diagram of treatment. 2.5 × 10 5 YUMMER1.7 cells were subcutaneously inoculated in C57BL/6 mice. When tumors reached ∼70 mm³ (days 8–10), mice were treated with anti-ICAM-1 or rat IgG isotype control (2.5 mg/kg, i.p.) every other day for seven doses; tumors were collected on days 21–24. (B-C) Tumor growth curves showing group means ± SEM (B) and individual trajectories (C) for IgG (black, n = 10) and anti-ICAM-1 (blue, n = 10) pooled from three independent experiments. P -value at the final time point by two-way ANOVA. (D) Representative immunofluorescence showing CD8⁺ T-cell distribution (green) across periphery to core in IgG- and anti-ICAM-1–treated tumors. Solid line marks the tumor boundary, and dashed line indicates the periphery–core boundary. Scale bar, 200 µm. (E) Quantification of CD8⁺ T-cell counts in periphery vs core (IgG, n = 6; anti-ICAM-1, n = 6). Statistical analysis was performed using unpaired or paired student two-tailed Student’s t-test. ( F-J ) Flow cytometric analysis of whole tumors comparing the frequency of CD8 + T cells among live cells (F), Granzyme B + cells among CD8 + T cells (G), and CD45 + immune cells among live cells (H), CD4 + T cells (I) among live cells, and regulatory T cells (Treg; CD4 + Foxp3 + ) among CD4 + T cells (J) between IgG (black, n = 14) and anti-ICAM-1 (blue, n =14). Statistical analysis was performed unpaired two-tailed Student’s t-test. Each point represents one mouse; bars show mean ± SEM.

Techniques: Control, Immunofluorescence, Two Tailed Test

(A) Schematic diagram of treatment. 5 × 10 5 YUMM1.7 cells were subcutaneously inoculated in C57BL/6 mice. When tumors reached ∼70mm³ (days 9–12), mice were intraperitoneally treated with rat IgG isotype control, anti-ICAM-1 (2.5 mg/kg), anti-PD-1 (5mg/kg), or combination (anti-ICAM-1 [2.5mg/kg] and anti-PD-1 [5mg/kg]) every other day for seven doses; tumors were collected on days 22-25. (B) Tumor growth curve showing group means ± SEM for IgG (black, n = 10), anti-ICAM-1 (blue, n = 10), anti-PD-1 (red, n = 10), and combined treatment (anti-ICAM-1 plus anti-PD-1; green, n = 10). P -value at the final time point by two-way ANOVA. (C-F) Flow cytometry analysis of whole tumors comparing frequency of CD8 + T cells among live cells (C) and proportions of activated CD8 + T cells with granzyme B + (D), CD62 - CD44 + (effector memory cells) (E), and CD69 + (F) among CD8 + T-cells (IgG, n = 14; anti-ICAM-1, n = 14; anti-PD-1, n =14; anti-ICAM-1 plus anti-PD-1, n = 14). For C-F, each point represents one mouse; bars show ± SEM. Statistical analysis was performed using one-way ANOVA.

Journal: bioRxiv

Article Title: Spatial polarization of endothelial ICAM-1 governs T-cell exclusion in melanoma

doi: 10.64898/2026.03.19.712709

Figure Lengend Snippet: (A) Schematic diagram of treatment. 5 × 10 5 YUMM1.7 cells were subcutaneously inoculated in C57BL/6 mice. When tumors reached ∼70mm³ (days 9–12), mice were intraperitoneally treated with rat IgG isotype control, anti-ICAM-1 (2.5 mg/kg), anti-PD-1 (5mg/kg), or combination (anti-ICAM-1 [2.5mg/kg] and anti-PD-1 [5mg/kg]) every other day for seven doses; tumors were collected on days 22-25. (B) Tumor growth curve showing group means ± SEM for IgG (black, n = 10), anti-ICAM-1 (blue, n = 10), anti-PD-1 (red, n = 10), and combined treatment (anti-ICAM-1 plus anti-PD-1; green, n = 10). P -value at the final time point by two-way ANOVA. (C-F) Flow cytometry analysis of whole tumors comparing frequency of CD8 + T cells among live cells (C) and proportions of activated CD8 + T cells with granzyme B + (D), CD62 - CD44 + (effector memory cells) (E), and CD69 + (F) among CD8 + T-cells (IgG, n = 14; anti-ICAM-1, n = 14; anti-PD-1, n =14; anti-ICAM-1 plus anti-PD-1, n = 14). For C-F, each point represents one mouse; bars show ± SEM. Statistical analysis was performed using one-way ANOVA.

Techniques: Control, Flow Cytometry

Expression levels of hyaluronidase (Hyal)-1 (A) , CD44 (B) and receptor for hyaluronan-mediated motility (RHAMM) (C) in the retinal lysates of non-diabetic control rats (C) (n=12) and diabetic rats (D) (n=12) were determined by Western blot analysis. After determination of the intensity of the protein bands, intensities were adjusted to those of β-actin in the samples. Oxidative stress was monitored with the use of 2’,7’-Dichlorofluorescein (DCF) fluorescence intensity analysis (D) . Results are expressed as mean ± standard deviation. Ultra-Low molecular weight hyaluronan (ULMW-HA) induces breakdown of blood-retinal barrier (E) . ULMW-HA was injected intravitreally at the dose of 50 ng in 5 µL in one eye and the same volume of phosphate-buffered saline (PBS) was injected in the contralateral eye of normal rats. The BRB was quantified with the fluorescein isothiocyanate-conjugated dextran technique. Results are expressed as mean ± standard deviation of 12 rats. *p < 0.05 compared to the values obtained from PBS-injected eyes. (independent t-test). Western blot analysis of retinas demonstrated that intravitreal injection of ULMW-HA induced significant upregulation of the expression of phospho-NF-κB (F) , phospho-ERK1/2 (G) , vascular endothelial growth factor (VEGF) (H) , intercellular adhesion molecule-1 (ICAM-1) (I) , vascular cell adhesion molecule-1 (VCAM-1) (J) and high-mobility group box-1 (HMGB1) (K) . Results are expressed as mean ± standard deviation or standard error of mean of 8–10 rats in each group (*p < 0.05; independent t-test).

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Expression levels of hyaluronidase (Hyal)-1 (A) , CD44 (B) and receptor for hyaluronan-mediated motility (RHAMM) (C) in the retinal lysates of non-diabetic control rats (C) (n=12) and diabetic rats (D) (n=12) were determined by Western blot analysis. After determination of the intensity of the protein bands, intensities were adjusted to those of β-actin in the samples. Oxidative stress was monitored with the use of 2’,7’-Dichlorofluorescein (DCF) fluorescence intensity analysis (D) . Results are expressed as mean ± standard deviation. Ultra-Low molecular weight hyaluronan (ULMW-HA) induces breakdown of blood-retinal barrier (E) . ULMW-HA was injected intravitreally at the dose of 50 ng in 5 µL in one eye and the same volume of phosphate-buffered saline (PBS) was injected in the contralateral eye of normal rats. The BRB was quantified with the fluorescein isothiocyanate-conjugated dextran technique. Results are expressed as mean ± standard deviation of 12 rats. *p < 0.05 compared to the values obtained from PBS-injected eyes. (independent t-test). Western blot analysis of retinas demonstrated that intravitreal injection of ULMW-HA induced significant upregulation of the expression of phospho-NF-κB (F) , phospho-ERK1/2 (G) , vascular endothelial growth factor (VEGF) (H) , intercellular adhesion molecule-1 (ICAM-1) (I) , vascular cell adhesion molecule-1 (VCAM-1) (J) and high-mobility group box-1 (HMGB1) (K) . Results are expressed as mean ± standard deviation or standard error of mean of 8–10 rats in each group (*p < 0.05; independent t-test).

Article Snippet: To determine the presence of Hyal-1, Hyal-2, HAS2, CD44, syndecan-1, heparan sulphate and RHAMM in the vitreous samples, equal volumes (10 μL) of vitreous samples were boiled in Laemmli’s sample buffer (1:1, v/v) under reducing condition for 10 min. Immunodetection was performed with the use of rabbit polyclonal anti-Hyal-1 antibody (1:1000, NBP2-16906, Novus Biologicals), mouse polyclonal anti-Hyal-2 antibody (1:1000, H00008692-B02P, Novus Biologicals), mouse monoclonal anti-HAS2 antibody (1:1000, ab140671, Abcam), rabbit monoclonal anti-CD44 antibody (1:1000, ab189524, Abcam), rabbit monoclonal anti-RHAMM antibody (1:1000, ab124729, Abcam), rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1000, MAB1018, R&D Systems), rabbit polyclonal anti-p65 subunit of nuclear factor-kappa B (phospho-NF-κB) (1:1000, NB100-82086, Novus Biologicals), rabbit polyclonal anti-high-mobility group box1 (HMGB1) (1:1000, Cat. no. ab18256, Abcam), mouse monoclonal anti-VEGF antibody (1:750, MAB293, R&D Systems), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:100, sc-8439, Santa Cruz Biotechnology Inc.), and mouse monoclonal anti-vascular cell adhesion molecule-1 (VCAM-1) antibody (1:100, sc-13160, Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Control, Western Blot, Fluorescence, Standard Deviation, Molecular Weight, Injection, Saline

Human retinal microvascular endothelial cells (HRMECs) were left untreated or stimulated with tumor necrosis factor-α (TNF-α) (5 ng/mL) for 24 h with or without apigenin (10 µg/mL). Protein expression of intercellular adhesion molecule-1 (ICAM-1) (A) and vascular cell adhesion molecule-1 (VCAM-1) (B) was determined by Western blot analysis. Adhesion of fluorescently labeled THP-1 monocytic cells to HRMECs monolayer was quantified (C) . Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells. #p < 0.05 compared with values obtained from cells treated with TNF-α (RFU = relative fluorescence unit).

Journal: Frontiers in Immunology

Article Title: Dysregulated hyaluronan metabolism drives inflammation and angiogenesis in proliferative diabetic retinopathy

doi: 10.3389/fimmu.2026.1724199

Figure Lengend Snippet: Human retinal microvascular endothelial cells (HRMECs) were left untreated or stimulated with tumor necrosis factor-α (TNF-α) (5 ng/mL) for 24 h with or without apigenin (10 µg/mL). Protein expression of intercellular adhesion molecule-1 (ICAM-1) (A) and vascular cell adhesion molecule-1 (VCAM-1) (B) was determined by Western blot analysis. Adhesion of fluorescently labeled THP-1 monocytic cells to HRMECs monolayer was quantified (C) . Results are expressed as mean ± standard deviation or standard error of mean from three different experiments each performed in triplicate. One-way ANOVA and independent t-test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from untreated cells. #p < 0.05 compared with values obtained from cells treated with TNF-α (RFU = relative fluorescence unit).

Techniques: Expressing, Western Blot, Labeling, Standard Deviation, Fluorescence

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