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Proteintech icam 1
Icam 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 401 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+icam+1/Icam-1+Antibody/pm41906653-153-13-15
Average 96 stars, based on 401 article reviews
icam 1 - by Bioz Stars, 2026-10
96/100 stars

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Related Articles

Incubation:

Article Title: Ultrasound assisted homing of human umbilical cord mesenchymal stem cells promotes recovery from acute respiratory distress syndrome.
Article Snippet: .. Proteins were electrophoretically transferred to PVDF membranes by wet transfer, closed for 5 min at room temperature using a protein-free rapid closure solution (Servicebio), and incubated overnight with the following antibodies: anti-SDF-1(1:1000, Abcam), anti-ICAM-1(1:2000, Proteintech), the membrane was exposed to horseradish peroxidase (HRP)-coupled goat anti-rabbit/anti-mouse IgG secondary antibody (1:10,000, Proteintech) for 1 h at room temperature. .. Finally, target bands were visualized using enhanced chemiluminescence reagents (Millipore), quantified using ImageJ software, and normalized according to the internal control GAPDH. qRT‐PCR To verify the changes in cytokine expression levels, mRNA expression of Sdf1, Icam1, Tnf, Il6, Il1b, Igf1 and Cxcl5 was detected using qRT-PCR.

Article Title: Tumor-Derived CCL16 Normalizes Tumor Vasculature through Macrophage ICAM-1 Receptor and Enhances Immunotherapy Efficacy in Hepatocellular Carcinoma
Article Snippet: .. Briefly, cell lysates containing 500 μg of total protein were incubated with 2 μg of anti-CCL16 (YN1319, Immunoway) or anti–ICAM-1 (10831-1-AP, Proteintech) overnight at 4°C. .. The immune complexes were captured using Protein A/G agarose beads (Invitrogen) for 2 hours at 4°C.

Article Title: Exosomes of Porphyromonas gingivalis-Infected Macrophages Impair the Endothelial Barrier and Angiogenesis In Vitro.
Article Snippet: Objectives: Endothelial dysfunction is a key contributor to periodontal disease and apical periodontitis.. However, the role of macrophages in mediating endothelial function via exosomes in these inflammatory diseases remains elusive.. Materials and Methods: Exosomes isolated from Porphyromonas gingivalis (P.g)infected THP1derived macrophages (P.gExos) and uninfected macrophages (ConExos) were verified and their effects on human umbilical vein endothelial cells (HUVECs) were investigated.

Article Title: Tumor-Derived CCL16 Normalizes Tumor Vasculature through Macrophage ICAM-1 Receptor and Enhances Immunotherapy Efficacy in Hepatocellular Carcinoma
Article Snippet: Equal amounts of protein (30 μg per lane) were separated by 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes (Millipore). .. The membranes were blocked with 5% non-fat milk in Tris-Buffered Saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: anti–β-tubulin (TA-10, ZsBio), anti–β-actin (TA-09, ZsBio), anti-CCL16 (YN1319, Immunoway), anti–ICAM-1 (10831-1-AP, Proteintech), anti-JAK2 (ET1607-35, HUABIO), anti–pJAK2 (ET1607-34, HUABIO), anti-STAT6 (51073-1-AP, Proteintech), anti–pSTAT6 (ab188080, Abcam), anti-DPP4 (DF12387, Affinity), anti-IL24 (26772-1-AP, Proteintech), anti-F4/80 (28463-1-AP, Proteintech), anti-CCR1 (DF2710, Affinity), anti-CD31 (ab124432, Abcam), anti-SMA (ZM-0003, ZsBio), anti-GST (#2625, Cell Signaling Technology), and anti-FLAG (F1804, Sigma-Aldrich). ..

Article Title: Ultrasound assisted homing of human umbilical cord mesenchymal stem cells promotes recovery from acute respiratory distress syndrome
Article Snippet: .. Proteins were electrophoretically transferred to PVDF membranes by wet transfer, closed for 5 min at room temperature using a protein-free rapid closure solution (Servicebio), and incubated overnight with the following antibodies: anti-SDF-1(1:1000, Abcam), anti-ICAM-1(1:2000, Proteintech), the membrane was exposed to horseradish peroxidase (HRP)-coupled goat anti-rabbit/anti-mouse IgG secondary antibody (1:10,000, Proteintech) for 1 h at room temperature. .. Finally, target bands were visualized using enhanced chemiluminescence reagents (Millipore), quantified using ImageJ software, and normalized according to the internal control GAPDH.

Article Title: Tumor-derived CCL16 Normalizes Tumor Vasculature through Macrophage ICAM-1 Receptor and Enhances Immunotherapy Efficacy in Hepatocellular Carcinoma
Article Snippet: Hepatocellular carcinoma (HCC) is characterized by aberrant tumor vasculature and an immunosuppressive tumor microenvironment (TME), both of which compromise immunotherapy efficacy while promoting circulating tumor cell (CTC) dissemination and immune escape.. In this study, we aimed to identify potential therapeutic targets for remodeling aberrant tumor vasculature by analyzing CTCs from patients with early-stage HCC.. HCC tissue samples derived from patients with elevated CTC counts demonstrated significant CCL16 downregulation accompanied by vascular structural abnormalities and an immunosuppressive TME.

Membrane:

Article Title: Ultrasound assisted homing of human umbilical cord mesenchymal stem cells promotes recovery from acute respiratory distress syndrome.
Article Snippet: .. Proteins were electrophoretically transferred to PVDF membranes by wet transfer, closed for 5 min at room temperature using a protein-free rapid closure solution (Servicebio), and incubated overnight with the following antibodies: anti-SDF-1(1:1000, Abcam), anti-ICAM-1(1:2000, Proteintech), the membrane was exposed to horseradish peroxidase (HRP)-coupled goat anti-rabbit/anti-mouse IgG secondary antibody (1:10,000, Proteintech) for 1 h at room temperature. .. Finally, target bands were visualized using enhanced chemiluminescence reagents (Millipore), quantified using ImageJ software, and normalized according to the internal control GAPDH. qRT‐PCR To verify the changes in cytokine expression levels, mRNA expression of Sdf1, Icam1, Tnf, Il6, Il1b, Igf1 and Cxcl5 was detected using qRT-PCR.

Article Title: Ultrasound assisted homing of human umbilical cord mesenchymal stem cells promotes recovery from acute respiratory distress syndrome
Article Snippet: .. Proteins were electrophoretically transferred to PVDF membranes by wet transfer, closed for 5 min at room temperature using a protein-free rapid closure solution (Servicebio), and incubated overnight with the following antibodies: anti-SDF-1(1:1000, Abcam), anti-ICAM-1(1:2000, Proteintech), the membrane was exposed to horseradish peroxidase (HRP)-coupled goat anti-rabbit/anti-mouse IgG secondary antibody (1:10,000, Proteintech) for 1 h at room temperature. .. Finally, target bands were visualized using enhanced chemiluminescence reagents (Millipore), quantified using ImageJ software, and normalized according to the internal control GAPDH.

other:

Article Title: ARF6 controls VSMC cell phenotypic switching upon lipid stimulation to promote inflammatory signaling contributing to the progression of atherosclerosis
Article Snippet: Anti-ICAM-1, anti-CD36 and anti-OLR1 were from Proteintech (Chicago, IL).

Article Title: ARF6 controls VSMC phenotypic switching upon lipid stimulation to promote inflammatory signaling contributing to the progression of atherosclerosis
Article Snippet: Anti-ARF1 (202260-1-ap), anti-ICAM-1 (10020-1-AP), anti-CD36 (18836-1-AP), anti-TLR4 (19811-1-ap), and anti-OLR1 (LOX-1) (11837-1-AP) were from Proteintech.

Saline:

Article Title: Tumor-Derived CCL16 Normalizes Tumor Vasculature through Macrophage ICAM-1 Receptor and Enhances Immunotherapy Efficacy in Hepatocellular Carcinoma
Article Snippet: Equal amounts of protein (30 μg per lane) were separated by 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes (Millipore). .. The membranes were blocked with 5% non-fat milk in Tris-Buffered Saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: anti–β-tubulin (TA-10, ZsBio), anti–β-actin (TA-09, ZsBio), anti-CCL16 (YN1319, Immunoway), anti–ICAM-1 (10831-1-AP, Proteintech), anti-JAK2 (ET1607-35, HUABIO), anti–pJAK2 (ET1607-34, HUABIO), anti-STAT6 (51073-1-AP, Proteintech), anti–pSTAT6 (ab188080, Abcam), anti-DPP4 (DF12387, Affinity), anti-IL24 (26772-1-AP, Proteintech), anti-F4/80 (28463-1-AP, Proteintech), anti-CCR1 (DF2710, Affinity), anti-CD31 (ab124432, Abcam), anti-SMA (ZM-0003, ZsBio), anti-GST (#2625, Cell Signaling Technology), and anti-FLAG (F1804, Sigma-Aldrich). ..



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(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 <t>of</t> <t>ICAM-1</t> (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.
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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