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Fig. 2 Pg W83 increases permeability of endothelial cell monolayers in a gingipain‐dependent manner. HDMEC (A–C) or HMEC‐1 (D) cells were grown to confluent monolayers on fibronectin‐coated inserts, infected with Pg at a MOI 1000 for 1.5 h, followed by the measurement of high molecular weight (70 kDa) fluorescein dextran passing through the monolayer barrier. Morphology of HDMEC monolayer in the absence (−Pg) or presence (+Pg) of W83 (A); white arrows indicate areas of cell attachment loss (scale bar in A = 20 μm). In vitro permeability assay of HDMEC in the absence (−Pg) or presence (+Pg) of W83 (B) and upon treatment with KYT inhibitors prior to HDMEC Pg infection (C). In vitro permeability assay of HMEC‐1 infected with W83 or ΔK/R‐ab (D). Data in B&C are presented as means ± SD and were analysed by one‐way ANOVA followed by Tukey’s post hoc comparison test. *P < 0.05, ***P < 0.001. Data in D are presented as means ± SD and were analysed by Student's t‐test. ***P < 0.001; for all experiments, n = 3 individual experiments with each individual experiment performed in triplicate technical repeats.