Extended Data Fig. 4: Syndecan-4 expression is required to promote vascular protection by ANGPTL4.
From: Angiopoietin-like 4 protects against endothelial dysfunction during bacterial sepsis
a-e, HDMEC were transfected with small interfering RNA (siRNA) against SDC4 or non-targeting control siRNA (CTL). 48 h post-transfection, cells were either uninfected or infected with N. meningitidis 2C4.3 for 30 min and treated with 1 μg.mL−1 of rhANGPTL4 or vehicle for 2 h and 30 min. a, SDC4 depletion efficiency was evaluated by immunoblotting using β-tubulin as a loading control. Bottom: shown are the ratio between SCD4 and β-tubulin signals. b, c, Cells were fixed and stained for VE-cadherin and DAPI and analysed by fluorescent microscopy (scale bars, 50 μm). d,e, Quantifications of VE-cadherin enrichment at intercellular junctions and of intercellular spaces area were performed with ImageJ software. Error bars show mean ± s.e.m. (n = 10 fields from n = 2 independent experiments); Unpaired t-test Welch’s correction (two-tailed) ****P < 0.001; NS, not significant (NS1 = 0.7432, NS2 = 3703, NS3 = 0.6109, NS4 = 0.538).
