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R&D Systems anti human tnfrsf10b antibody
Nanoparticle flowcytometry analysis of <t>TNFRSF10B/CD63</t> dual labelling extracellular vesicles ( EVs) . ( a) Scatter plots display populations of EVs collected at four different experimental conditions. Each scatter plot is convened into four quarters based on baseline cutoffs on the green or red fluorescent signals. Q1 for TNFRSF10B+ only, Q2 for both TNFRSF10B+ and CD63+, Q3 for CD63+ only, and Q4 for non‐specific nano particles, with the numbers show corresponding percentages of each quarter. ( b) The bar graph compares EV populations, with the height of each bar indicating the percentage for individual EV population (P Q1 , P Q2 and P Q3 ). Samples from the 2nd, 3rd and 4th collections were indicated in blue, orange and grey colours, accordingly ( p < 0.05). P Q1 , P Q2 and P Q3 were the average of the corresponding collections from the 2nd and the 3rd round. Error bars show standard deviation.
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Bio-Techne corporation antibody against human trail r2
A-B) Percentage of <t>TRAIL-R1</t> and <t>TRAIL-R2</t> expression on PMN-MDSCs and M-MDSCs in HIV-1 infected mice (2 donors, n=3-4 per group). C) t-SNE analysis of mCD45 - hCD45 + TRAIL + cells in HIV-1 infected mice (1 donor, n=4 mice per donor) overlayed with expression heatmaps of CD56, CD3 and TRAIL. D) Percentage of TRAIL expressing NK cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). E) Frequency of TRAIL expressing T cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05, **P < 0.01, **P < 0.005 and ****P < 0.0001.
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R&D Systems alexa488 antihuman tnfrsf10b antibody
A-B) Percentage of <t>TRAIL-R1</t> and <t>TRAIL-R2</t> expression on PMN-MDSCs and M-MDSCs in HIV-1 infected mice (2 donors, n=3-4 per group). C) t-SNE analysis of mCD45 - hCD45 + TRAIL + cells in HIV-1 infected mice (1 donor, n=4 mice per donor) overlayed with expression heatmaps of CD56, CD3 and TRAIL. D) Percentage of TRAIL expressing NK cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). E) Frequency of TRAIL expressing T cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05, **P < 0.01, **P < 0.005 and ****P < 0.0001.
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R&D Systems anti human pe conjugated antibody against trail r2
Nutlin-3a-loaded ethosomes upregulate surface death receptors <t>TRAIL-R2</t> of p53 wild-type melanoma cells. Surface cytofluorimetric analysis of HT144 expressing p53 wild-type ( A ) and SK-MEL-28 expressing p53 mut ( B ) cell cultures treated with nutlin-3a or nutlin-3a-loaded ethosomes with equivalent nutlin-3a concentrations after 24 h. Vehicle and empty ethosomes are reported as control vehicles. Data are calculated as the fold increase in median fluorescence values with respect to untreated cells (set to 1). Bars represent the mean ± SEM, and each circle denotes the value of each experimental replicate. Statistical analysis was performed by means of ANOVA followed by Bonferroni’s post hoc test. *** indicates a p -value ≤ 0.001.
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Novus Biologicals anti human trail r2 mab
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
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ACROBiosystems recombinant human trail r2
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
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ATCC control trail r2
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
Control Trail R2, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ATCC trail r2 treatment
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
Trail R2 Treatment, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ATCC trail r2 fc
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
Trail R2 Fc, supplied by ATCC, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Kamiya nkg2d tnbc promote apoptosis microrna 519a 3p trail r2 caspase 7
( A ) <t>Trail</t> , Trail-R1 , and <t>Trail-R2</t> mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.
Nkg2d Tnbc Promote Apoptosis Microrna 519a 3p Trail R2 Caspase 7, supplied by Kamiya, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Nanoparticle flowcytometry analysis of TNFRSF10B/CD63 dual labelling extracellular vesicles ( EVs) . ( a) Scatter plots display populations of EVs collected at four different experimental conditions. Each scatter plot is convened into four quarters based on baseline cutoffs on the green or red fluorescent signals. Q1 for TNFRSF10B+ only, Q2 for both TNFRSF10B+ and CD63+, Q3 for CD63+ only, and Q4 for non‐specific nano particles, with the numbers show corresponding percentages of each quarter. ( b) The bar graph compares EV populations, with the height of each bar indicating the percentage for individual EV population (P Q1 , P Q2 and P Q3 ). Samples from the 2nd, 3rd and 4th collections were indicated in blue, orange and grey colours, accordingly ( p < 0.05). P Q1 , P Q2 and P Q3 were the average of the corresponding collections from the 2nd and the 3rd round. Error bars show standard deviation.

Journal: Journal of Extracellular Biology

Article Title: Effects of electric fields on the release and content of extracellular vesicles

doi: 10.1002/jex2.70018

Figure Lengend Snippet: Nanoparticle flowcytometry analysis of TNFRSF10B/CD63 dual labelling extracellular vesicles ( EVs) . ( a) Scatter plots display populations of EVs collected at four different experimental conditions. Each scatter plot is convened into four quarters based on baseline cutoffs on the green or red fluorescent signals. Q1 for TNFRSF10B+ only, Q2 for both TNFRSF10B+ and CD63+, Q3 for CD63+ only, and Q4 for non‐specific nano particles, with the numbers show corresponding percentages of each quarter. ( b) The bar graph compares EV populations, with the height of each bar indicating the percentage for individual EV population (P Q1 , P Q2 and P Q3 ). Samples from the 2nd, 3rd and 4th collections were indicated in blue, orange and grey colours, accordingly ( p < 0.05). P Q1 , P Q2 and P Q3 were the average of the corresponding collections from the 2nd and the 3rd round. Error bars show standard deviation.

Article Snippet: In detail, 100 µL of isolated EV sample was mixed with 4 µL of Alexa488 anti‐human TNFRSF10B antibody (FAB6311G, R&D Systems, Minneapolis, MN, USA) and 2 µL of PE anti‐human CD63 antibody (561925, BD Biosciences, Franklin Lakes, NJ, USA) and incubated at 37°C for 30 min. Unlabelled antibodies were removed using a second application of SEC with a fresh qEV single column (70 nm Gen 2, IZON, Medford, MA).

Techniques: Standard Deviation

A-B) Percentage of TRAIL-R1 and TRAIL-R2 expression on PMN-MDSCs and M-MDSCs in HIV-1 infected mice (2 donors, n=3-4 per group). C) t-SNE analysis of mCD45 - hCD45 + TRAIL + cells in HIV-1 infected mice (1 donor, n=4 mice per donor) overlayed with expression heatmaps of CD56, CD3 and TRAIL. D) Percentage of TRAIL expressing NK cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). E) Frequency of TRAIL expressing T cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05, **P < 0.01, **P < 0.005 and ****P < 0.0001.

Journal: bioRxiv

Article Title: TRAIL-mediated PMN-MDSC depletion prevents CD4 + T cell loss in HIV-infected humanized mice

doi: 10.1101/2025.01.22.634356

Figure Lengend Snippet: A-B) Percentage of TRAIL-R1 and TRAIL-R2 expression on PMN-MDSCs and M-MDSCs in HIV-1 infected mice (2 donors, n=3-4 per group). C) t-SNE analysis of mCD45 - hCD45 + TRAIL + cells in HIV-1 infected mice (1 donor, n=4 mice per donor) overlayed with expression heatmaps of CD56, CD3 and TRAIL. D) Percentage of TRAIL expressing NK cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). E) Frequency of TRAIL expressing T cells in blood, liver and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post intraperitoneal infection with Q23.17 (10 5 IU) and uninfected human donor-matched controls. (3 donors, n=3-4 mice per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05, **P < 0.01, **P < 0.005 and ****P < 0.0001.

Article Snippet: To deplete human MDSCs, mice received 100 µg of antibody against human TRAIL-R2 (MAB631; Bio-Techne R&D Systems, Minneapolis, MN, USA) by i.p. injection in a volume of 200 µl starting 3 days post-infection and then weekly for 8 weeks.

Techniques: Expressing, Infection

A, B) Percentage and Total numbers of PMN-MDSCs in the livers and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies. Data is presented from individual mice from 2-4 human donor-matched cohorts represented by matching symbols (n=3-5 per donor, per experimental group). C, D) Percentage and Total numbers of M-MDSCs in the livers and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies. Data is presented from individual mice from 2-4 human donor-matched cohorts represented by matching symbols (n=3-5 per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05. E, F) Plasma viral RNA load of Hu-NSG-Tg(IL-15) mice infected with Q23.17 and treated with anti-TRAIL-R2 or untreated donor-matched control mice. Data from each individual mouse is depicted by a different symbol (2 donors, n=4-5 per experimental group, per donor). G, H) Percentage of CD4+ T cells of total T cells in the peripheral blood before and after HIV-1 infection in anti-TRAIL-R2 treated and untreated donor matched Hu-NSG-Tg(IL-15) mice. Data from each individual mouse is depicted by a different symbol (2 donors, n=4-5 per experimental group, per donor). Statistical significance was calculated using paired t-tests. *P < 0.05.

Journal: bioRxiv

Article Title: TRAIL-mediated PMN-MDSC depletion prevents CD4 + T cell loss in HIV-infected humanized mice

doi: 10.1101/2025.01.22.634356

Figure Lengend Snippet: A, B) Percentage and Total numbers of PMN-MDSCs in the livers and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies. Data is presented from individual mice from 2-4 human donor-matched cohorts represented by matching symbols (n=3-5 per donor, per experimental group). C, D) Percentage and Total numbers of M-MDSCs in the livers and spleens of Hu-NSG-Tg(IL-15) mice 8 weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies. Data is presented from individual mice from 2-4 human donor-matched cohorts represented by matching symbols (n=3-5 per donor, per experimental group). Statistical significance was calculated using unpaired t-tests. *P < 0.05. E, F) Plasma viral RNA load of Hu-NSG-Tg(IL-15) mice infected with Q23.17 and treated with anti-TRAIL-R2 or untreated donor-matched control mice. Data from each individual mouse is depicted by a different symbol (2 donors, n=4-5 per experimental group, per donor). G, H) Percentage of CD4+ T cells of total T cells in the peripheral blood before and after HIV-1 infection in anti-TRAIL-R2 treated and untreated donor matched Hu-NSG-Tg(IL-15) mice. Data from each individual mouse is depicted by a different symbol (2 donors, n=4-5 per experimental group, per donor). Statistical significance was calculated using paired t-tests. *P < 0.05.

Article Snippet: To deplete human MDSCs, mice received 100 µg of antibody against human TRAIL-R2 (MAB631; Bio-Techne R&D Systems, Minneapolis, MN, USA) by i.p. injection in a volume of 200 µl starting 3 days post-infection and then weekly for 8 weeks.

Techniques: Infection, Control

Percentage of cells in each cluster generated from unsupervised FlowSOM clustering. A-C) t-SNE dimensionality reduction of hCD45+mCD45-cells positive for either CD3 and/or CD56 with overlayed with FlowSOM clusters of splenic immune cells Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched control mice. Cells from individual mice were pooled and downsampled to 50,000 events prior to t-SNE and FlowSOM analysis.

Journal: bioRxiv

Article Title: TRAIL-mediated PMN-MDSC depletion prevents CD4 + T cell loss in HIV-infected humanized mice

doi: 10.1101/2025.01.22.634356

Figure Lengend Snippet: Percentage of cells in each cluster generated from unsupervised FlowSOM clustering. A-C) t-SNE dimensionality reduction of hCD45+mCD45-cells positive for either CD3 and/or CD56 with overlayed with FlowSOM clusters of splenic immune cells Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched control mice. Cells from individual mice were pooled and downsampled to 50,000 events prior to t-SNE and FlowSOM analysis.

Article Snippet: To deplete human MDSCs, mice received 100 µg of antibody against human TRAIL-R2 (MAB631; Bio-Techne R&D Systems, Minneapolis, MN, USA) by i.p. injection in a volume of 200 µl starting 3 days post-infection and then weekly for 8 weeks.

Techniques: Generated, Infection, Control

A) t-SNE dimensionality reduction of hCD45+mCD45-cells positive for either CD3 and/or CD56 with overlayed with FlowSOM clusters of splenic immune cells Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched control mice. Cells from individual mice were pooled and downsampled to 50,000 events prior to t-SNE and FlowSOM analysis B) Percentage of cells in Cluster 3 (NK Cells) generated from unsupervised FlowSOM clustering. C) Heatmap displaying phenotypic marker expression for each population clustered using FlowSOM algorithm. D-E) Extracellular expression of NKG2D and CD107a (Lamp-1) on NK cells (CD56 + CD3 - ) and CTLs (CD56 - CD3 + CD8 + ) in Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched controls (2 donors, n=3-5 mice per donor, per experimental group). F-G) Intracellular expression of TNFa and intranucleuar expression of Ki67 on NK cells (CD56 + CD3 - ) and CTLs (CD56 - CD3 + CD8 + ) in Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched controls (2 donors, n=3-5 mice per donor, per experimental group). Statistical significance was calculated using an unpaired t-test *P < 0.05, **P < 0.01, **P < 0.005, and ****P < 0.0001.

Journal: bioRxiv

Article Title: TRAIL-mediated PMN-MDSC depletion prevents CD4 + T cell loss in HIV-infected humanized mice

doi: 10.1101/2025.01.22.634356

Figure Lengend Snippet: A) t-SNE dimensionality reduction of hCD45+mCD45-cells positive for either CD3 and/or CD56 with overlayed with FlowSOM clusters of splenic immune cells Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched control mice. Cells from individual mice were pooled and downsampled to 50,000 events prior to t-SNE and FlowSOM analysis B) Percentage of cells in Cluster 3 (NK Cells) generated from unsupervised FlowSOM clustering. C) Heatmap displaying phenotypic marker expression for each population clustered using FlowSOM algorithm. D-E) Extracellular expression of NKG2D and CD107a (Lamp-1) on NK cells (CD56 + CD3 - ) and CTLs (CD56 - CD3 + CD8 + ) in Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched controls (2 donors, n=3-5 mice per donor, per experimental group). F-G) Intracellular expression of TNFa and intranucleuar expression of Ki67 on NK cells (CD56 + CD3 - ) and CTLs (CD56 - CD3 + CD8 + ) in Hu-NSG-Tg(IL-15) mice 8-weeks post HIV-1 infection, with or without treatment with anti-TRAIL-R2 agonistic antibodies and uninfected donor-matched controls (2 donors, n=3-5 mice per donor, per experimental group). Statistical significance was calculated using an unpaired t-test *P < 0.05, **P < 0.01, **P < 0.005, and ****P < 0.0001.

Article Snippet: To deplete human MDSCs, mice received 100 µg of antibody against human TRAIL-R2 (MAB631; Bio-Techne R&D Systems, Minneapolis, MN, USA) by i.p. injection in a volume of 200 µl starting 3 days post-infection and then weekly for 8 weeks.

Techniques: Infection, Control, Generated, Marker, Expressing

Nutlin-3a-loaded ethosomes upregulate surface death receptors TRAIL-R2 of p53 wild-type melanoma cells. Surface cytofluorimetric analysis of HT144 expressing p53 wild-type ( A ) and SK-MEL-28 expressing p53 mut ( B ) cell cultures treated with nutlin-3a or nutlin-3a-loaded ethosomes with equivalent nutlin-3a concentrations after 24 h. Vehicle and empty ethosomes are reported as control vehicles. Data are calculated as the fold increase in median fluorescence values with respect to untreated cells (set to 1). Bars represent the mean ± SEM, and each circle denotes the value of each experimental replicate. Statistical analysis was performed by means of ANOVA followed by Bonferroni’s post hoc test. *** indicates a p -value ≤ 0.001.

Journal: Cells

Article Title: Enhanced Anti-Melanoma Activity of Nutlin-3a Delivered via Ethosomes: Targeting p53-Mediated Apoptosis in HT144 Cells

doi: 10.3390/cells13201678

Figure Lengend Snippet: Nutlin-3a-loaded ethosomes upregulate surface death receptors TRAIL-R2 of p53 wild-type melanoma cells. Surface cytofluorimetric analysis of HT144 expressing p53 wild-type ( A ) and SK-MEL-28 expressing p53 mut ( B ) cell cultures treated with nutlin-3a or nutlin-3a-loaded ethosomes with equivalent nutlin-3a concentrations after 24 h. Vehicle and empty ethosomes are reported as control vehicles. Data are calculated as the fold increase in median fluorescence values with respect to untreated cells (set to 1). Bars represent the mean ± SEM, and each circle denotes the value of each experimental replicate. Statistical analysis was performed by means of ANOVA followed by Bonferroni’s post hoc test. *** indicates a p -value ≤ 0.001.

Article Snippet: Cells were treated for 48 h with nutlin-3a in solution, nutlin-3a-loaded ethosomes, and the vehicle and controls as described above, harvested, and then stained with anti-human PE-conjugated antibody against TRAIL-R2 (clone 71908, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Control, Fluorescence

( A ) Trail , Trail-R1 , and Trail-R2 mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.

Journal: Science Advances

Article Title: The generation of stable microvessels in ischemia is mediated by endothelial cell derived TRAIL

doi: 10.1126/sciadv.adn8760

Figure Lengend Snippet: ( A ) Trail , Trail-R1 , and Trail-R2 mRNA expression in HMEC-1 after exposure to hypoxia (2% O 2 ) or normoxia (21% O 2 ) for 24 hours, qPCR normalized to β -actin ( n = 6 to 8 per treatment). ( B ) Total internal reflection fluorescence microscopy images (left) and quantification (right) of TRAIL cell surface expression ( n = 3 per treatment). ( C ) TRAIL-R2 cell surface expression by flow cytometry ( n = 4 per treatment). ( D ) Left: Western blot of TRAIL-R1 and β-actin. Right: TRAIL-R1 protein expression normalized to β-actin ( n = 4 per group). ( E ) Protein-protein interactions between TRAIL–TRAIL-R1 and TRAIL–TRAIL-R2, measured using the Duolink Proximity Ligation Assay. Interactions are indicated by red staining. Left: Representative image. Right: Quantification ( n = 6 per treatment). Results are means ± SEM; Students t test or two-way analysis of variance (ANOVA); * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: HMEC-1 were blocked in 10% bovine serum albumin and then stained with anti-human TRAIL-R2 mAb (20 μg/ml; Novus Biological) or IgG (Life Technologies) and anti-rabbit fluorescein isothiocyanate (FITC;1:500, BD Biosciences) for 45 min at 4°C.

Techniques: Expressing, Fluorescence, Microscopy, Flow Cytometry, Western Blot, Proximity Ligation Assay, Staining

Plasma ( A ) TRAIL and ( B ) TRAIL-R2 levels in PAD versus healthy individuals ( n = 11 to 12 per group). ( C ) Schematic illustrating tissue harvest locations from below-knee amputations. ( D ) Stable microvessel numbers in nonischemic and ischemic regions of amputated tissues. Left: Representative image of stable microvessels (CD31 + SMA + , yellow arrows). Laminin (myocytes, white), CD31 (ECs, red), and SMA (pericytes, green); scale bars, 20 μm. Middle: Microvessel quantification. Right: Myocyte number is reduced with ischemia ( n = 3 per group). ( E ) Myography showing impaired endothelial function in arteries isolated from nonischemic versus ischemic regions. Left: Arterial relaxation in response to increasing doses of acetylcholine (ACh). Right: No change in sodium nitroprusside (SNP)–mediated relaxation ( n = 5 per group). ( F ) Trail and ( G ) Nox4 mRNA expression in patient tissues ( n = 5 per group). mRNA normalized to β -actin . Results are means ± SEM; Mann-Whitney U test, paired t test, or two-way ANOVA; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Journal: Science Advances

Article Title: The generation of stable microvessels in ischemia is mediated by endothelial cell derived TRAIL

doi: 10.1126/sciadv.adn8760

Figure Lengend Snippet: Plasma ( A ) TRAIL and ( B ) TRAIL-R2 levels in PAD versus healthy individuals ( n = 11 to 12 per group). ( C ) Schematic illustrating tissue harvest locations from below-knee amputations. ( D ) Stable microvessel numbers in nonischemic and ischemic regions of amputated tissues. Left: Representative image of stable microvessels (CD31 + SMA + , yellow arrows). Laminin (myocytes, white), CD31 (ECs, red), and SMA (pericytes, green); scale bars, 20 μm. Middle: Microvessel quantification. Right: Myocyte number is reduced with ischemia ( n = 3 per group). ( E ) Myography showing impaired endothelial function in arteries isolated from nonischemic versus ischemic regions. Left: Arterial relaxation in response to increasing doses of acetylcholine (ACh). Right: No change in sodium nitroprusside (SNP)–mediated relaxation ( n = 5 per group). ( F ) Trail and ( G ) Nox4 mRNA expression in patient tissues ( n = 5 per group). mRNA normalized to β -actin . Results are means ± SEM; Mann-Whitney U test, paired t test, or two-way ANOVA; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: HMEC-1 were blocked in 10% bovine serum albumin and then stained with anti-human TRAIL-R2 mAb (20 μg/ml; Novus Biological) or IgG (Life Technologies) and anti-rabbit fluorescein isothiocyanate (FITC;1:500, BD Biosciences) for 45 min at 4°C.

Techniques: Isolation, Expressing, MANN-WHITNEY

( A ) Trail +/+ and Trail −/− EC proliferation is increased with MD5-1 at 24 hours, normalized to immunoglobulin G (IgG; n = 4 to 5 per group). ( B ) Representative images showing increased tubule formation of Trail +/+ ECs with MD5-1 versus IgG at 6 hours (10 ng/ml); scale bars, 500 μm. ( C ) Aortic rings from wild-type mice were exposed to MD5-1 or IgG for 4 days, and sprout number and total sprout length were assessed ( n = 3 to 4 per group). ( D ) Aortic rings from Trail-R −/− mice exposed to hypoxia for 6 days have reduced sprout number and reduced total sprout length ( n = 4 per group). Human TRAIL-R2 agonistic antibody (αTRAIL-R2, 10 ng/ml) stimulates HMEC-1 ( E ) proliferation, ( F ) migration, and ( G ) tubule formation ( n = 5 to 7 per group). Conversely, neutralization by αTRAIL-R1 has no effect on TRAIL-inducible HMEC-1 ( H ) proliferation and ( I ) migration ( n = 3 to 4 per group). Results are means ± SEM; two-way ANOVA, Mann-Whitney U test, or Students t test; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Journal: Science Advances

Article Title: The generation of stable microvessels in ischemia is mediated by endothelial cell derived TRAIL

doi: 10.1126/sciadv.adn8760

Figure Lengend Snippet: ( A ) Trail +/+ and Trail −/− EC proliferation is increased with MD5-1 at 24 hours, normalized to immunoglobulin G (IgG; n = 4 to 5 per group). ( B ) Representative images showing increased tubule formation of Trail +/+ ECs with MD5-1 versus IgG at 6 hours (10 ng/ml); scale bars, 500 μm. ( C ) Aortic rings from wild-type mice were exposed to MD5-1 or IgG for 4 days, and sprout number and total sprout length were assessed ( n = 3 to 4 per group). ( D ) Aortic rings from Trail-R −/− mice exposed to hypoxia for 6 days have reduced sprout number and reduced total sprout length ( n = 4 per group). Human TRAIL-R2 agonistic antibody (αTRAIL-R2, 10 ng/ml) stimulates HMEC-1 ( E ) proliferation, ( F ) migration, and ( G ) tubule formation ( n = 5 to 7 per group). Conversely, neutralization by αTRAIL-R1 has no effect on TRAIL-inducible HMEC-1 ( H ) proliferation and ( I ) migration ( n = 3 to 4 per group). Results are means ± SEM; two-way ANOVA, Mann-Whitney U test, or Students t test; * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: HMEC-1 were blocked in 10% bovine serum albumin and then stained with anti-human TRAIL-R2 mAb (20 μg/ml; Novus Biological) or IgG (Life Technologies) and anti-rabbit fluorescein isothiocyanate (FITC;1:500, BD Biosciences) for 45 min at 4°C.

Techniques: Migration, Neutralization, MANN-WHITNEY

(i) ECs are a major source of TRAIL in the circulation. (ii) TRAIL binds and activates TRAIL-R2 to stimulate the formation of immature microvessels. (iii) EC-derived TRAIL recruits pericytes to the endothelium to form stable blood vessel networks. EC-pericyte communication is mediated by HBEGF and HBEGF receptor interactions. (iv) EC-derived TRAIL is suppressed in PAD. (v) Agonistic TRAIL-R2 mAb stimulates the formation of stable microvessels in PAD.

Journal: Science Advances

Article Title: The generation of stable microvessels in ischemia is mediated by endothelial cell derived TRAIL

doi: 10.1126/sciadv.adn8760

Figure Lengend Snippet: (i) ECs are a major source of TRAIL in the circulation. (ii) TRAIL binds and activates TRAIL-R2 to stimulate the formation of immature microvessels. (iii) EC-derived TRAIL recruits pericytes to the endothelium to form stable blood vessel networks. EC-pericyte communication is mediated by HBEGF and HBEGF receptor interactions. (iv) EC-derived TRAIL is suppressed in PAD. (v) Agonistic TRAIL-R2 mAb stimulates the formation of stable microvessels in PAD.

Article Snippet: HMEC-1 were blocked in 10% bovine serum albumin and then stained with anti-human TRAIL-R2 mAb (20 μg/ml; Novus Biological) or IgG (Life Technologies) and anti-rabbit fluorescein isothiocyanate (FITC;1:500, BD Biosciences) for 45 min at 4°C.

Techniques: Derivative Assay