Fig. 3: Soluble HFE2 and RGMa effects on endothelial cells.
From: The liver and muscle secreted HFE2-protein maintains central nervous system blood vessel integrity
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, Hfe2ΔAlb-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; Hfe2fl/fl n = 3 and Hfe2ΔAlb-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).
