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Miltenyi Biotec mouse
Mouse, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cd54+antibody/CD54+(ICAM-1)+Antibody%2C+anti-mouse%2C+REAfinity/pmc13280562-439-17-22
Average 93 stars, based on 5 article reviews

mouse - by Bioz Stars, 2026-09

93/100 stars

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Article Title: Preclinical quality, safety, and efficacy of a CGMP iPSC-derived myogenic progenitor product for the treatment of muscular dystrophies.
Article Snippet: Karim Azzag,1,13 Alessandro Magli,1,2,13 James Kiley,1 Darin Sumstad,3 Diane Kadidlo,3 Sarah B. Crist,1 Beverly Norris,4 Aaron Ahlquist,1 Laura L. Hocum Stone,5 John Everett,6 Jill Shappa Faustich,5 Davis Seelig,7,8 Parthasarathy Rangarajan,5 Hyunkee Kim,1 Craig Flory,4 Frederic Bushman,6 Sabarinathan Ramachandran,2,5 Peter B. Kang,9,10 Robert J. Schumacher,4 John E. Wagner,2,11 Michael Kyba,2,9,11 Melanie L. Graham,5,12 David H. McKenna,2,3 and Rita C.R.. Perlingeiro1,2,9

Incubation:

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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 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

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