Extended Data Fig. 6: Antibody neutralisation of ANGPTL4 or SDC4 in primary human brain microvascular endothelial cells promotes vascular alteration and inflammation in response to meningococcal infection.
From: Angiopoietin-like 4 protects against endothelial dysfunction during bacterial sepsis
a-c, HBMEC grown on cell filters were either uninfected or infected with N. meningitidis 2C4.3 for 30 min and treated with 10 μg.mL−1 of anti-ANGPTL4 or anti-SDC4 antibodies or vehicle for 2h30. a, Transendothelial electrical resistance (TEER) was measured. Shown are the result of one representative experiment of two independent experiments performed in triplicate. Error bars show mean ± s.e.m; Ordinary one-way ANOVA Tukey’s multiple comparison test **P < 0.001; *P < 0.05. b, Cells were fixed and stained for VE-cadherin, Bacteria and DAPI and analysed by fluorescent microscopy. Representative images of 3 independent experiments are shown. c, Production of human cytokines (TNF, IL-6 and IL-8) and markers of vascular inflammation (E-selectin, ICAM-1, VCAM-1) or vascular alteration (Endoglin, Thrombomodulin) in the cell supernatants was quantified with multiplex electrochemiluminescent immunoassays. Shown are the result of one representative experiment performed in triplicate. Error bars show mean ± s.e.m; One-way ANOVA Dunnett’s multiple comparison test ****P < 0.0001; **P < 0.01; *P < 0.05; NS, not significant (NS1 = 0.9449, NS2 = 0.5652, NS3 = 0.3368, NS4 = 0.3843, NS5 = 0.6827, NS6 = 0.2803).
