murine mouse brain endothelial cell line bend 3 Search Results


86
Procell Inc murine cerebral microvascular endothelial cell line
Murine Cerebral Microvascular Endothelial Cell Line, supplied by Procell Inc, 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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99
ATCC murine brain derived endothelial cell line b end3
Murine Brain Derived Endothelial Cell Line B End3, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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96
ATCC murine brain endothelial cells bend3
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Murine Brain Endothelial Cells Bend3, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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95
ATCC cortical endothelioma cells
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Cortical Endothelioma Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
ATCC plasmid construction murine brain endothelial cells
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Plasmid Construction Murine Brain Endothelial Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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90
iCell Bioscience Inc murine microvascular endothelial cell line bend.3
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Murine Microvascular Endothelial Cell Line Bend.3, supplied by iCell Bioscience Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
Malvern Panalytical zetasizer advance
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Zetasizer Advance, supplied by Malvern Panalytical, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Micromeritics Instrument zetasizer advance
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
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NSJ Bioreagents thyroglobulin antibody
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Thyroglobulin Antibody, supplied by NSJ Bioreagents, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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95
ATCC aeromonas salmonicida subsp.achromogenes schubert
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Aeromonas Salmonicida Subsp.Achromogenes Schubert, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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99
ATCC nctc clone 929
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
Nctc Clone 929, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ATCC u-87 mg
a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in <t>bEnd3</t> cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).
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a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in bEnd3 cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).

Journal: Nature Communications

Article Title: The liver and muscle secreted HFE2-protein maintains central nervous system blood vessel integrity

doi: 10.1038/s41467-024-45303-1

Figure Lengend Snippet: a Knockout of liver Hfe2 reduces pericyte coverage in section from the cerebral cortex (mean ± s.e.m.; unpaired two-tail t-test; AAV8-GFP n = 3, Hfe2 ΔAlb-cre n = 4, and AAV8-AlbCre n = 3). Scale bar, 50μm. b Quantification through quantitative RT-PCR shows a reduction in PDGF-B mRNA levels in liver knock out animals (mean ± s.e.m.; paired two-tail t-test; Hfe2 fl/fl n = 3 and Hfe2 ΔAlb-cre n = 3). c Quantification of PDGF-B mRNA in cultured bEnd.3 cells following indicated treatments. Treatment with RGMa leads to a significant reduction of PDGF-B mRNA levels that is rescued with HFE2 addition (mean ± s.e.m.; paired two-tail t-test; HFE2 n = 3, RGMa n = 4, and RGMa+ HFE2 n = 5; n representing an independent experiment). d Immunocytochemistry of claudin-5 in human primary endothelial cell monolayer after PBS, HFE2, RGMa, and RGMa+HFE2 treatments and quantification (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). Scale bar, 100 µm. e Western blotting of Claudin-5 expression in bEnd3 cell lysates after indicated treatment (mean ± s.e.m.; unpaired two-tailed t-test; replicates n = 3). f Transwell permeability leakage assay performed on a monolayer of bEnd3 cells using HRP (mean ± s.e.m.; paired one-tail t-test; replicates n = 3). g Transwell permeability leakage assay performed on a monolayer of human primary endothelial cells using 70 kDa FITC-dextran (mean ± s.e.m.; paired two-tail t-test; replicates n = 3). h In vitro TEER analysis shows that RGMa alters the integrity of a monolayer of bEnd3 cells, this is rescued by HFE2 (mean ± s.e.m.; unpaired two-tail t-test; replicates n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. Source data are provided as a Source Data file (includes exact p-values).

Article Snippet: The endothelial monolayer permeability assay was performed as previously described DiStefano et al. Murine Brain Endothelial cells (bEnd3) (ATCC®CRL-2299 TM ) were seeded on 3 μm pore Transwell inserts (Corning Life Sciences) coated with 10 μg/ml human plasma fibronectin (Sigma, FC010).

Techniques: Knock-Out, Quantitative RT-PCR, Cell Culture, Immunocytochemistry, Western Blot, Expressing, Two Tailed Test, Permeability, In Vitro