Figure 1: Depletion of endothelial anxA8 impairs leukocyte rolling and adhesion. | Nature Communications

Figure 1: Depletion of endothelial anxA8 impairs leukocyte rolling and adhesion.

From: Annexin A8 controls leukocyte recruitment to activated endothelial cells via cell surface delivery of CD63

Figure 1

(a) Forty-eight hours after transfecting HUVEC with siRNA specific for anxA8 (siA8) or non-targeting siRNA (siCt), total cell lysates were prepared and analysed by immunoblotting using anti-anxA8 antibodies. α-Tubulin served as loading control (left panel). The right panel shows confluent HUVEC that were subjected to the same transfection regime and then placed in a parallel plate flow chamber and incubated with basal media or histamine-containing stimulation media for 20 min. The chambers were then perfused with isolated PMN for 7 min, while recording movies. The images shown represent the last section of the movies after histamine treatment of control and anxA8-depleted cells. Arrows mark firmly interacting leukocytes on the endothelial monolayers. Scale bars, 100 μm. Adhesion (b), rolling (c) and corresponding rolling velocities (d) of PMN on HUVEC monolayers of non-transfected (NT), siCt- or siA8-transfected cells were quantified and statistically analysed by one-way ANOVA followed by Tukey’s multiple comparison test. ***P<0.001. Data represent means±s.e.m. from at least 12 optical fields (b,c) or 31 rolling cells (d) from four independent experiments; AU, arbitrary units; NS, not significant.

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