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. 2019 May 15;10:1092. doi: 10.3389/fimmu.2019.01092

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Copyright © 2019 Tobia, Chiodelli, Barbieri, Buraschi, Ferrari, Mitola, Borsani, Guerra and Presta.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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ACKR3 generates a chemotactic CXCL12 gradient for HUVECs. (A) Schematic representation of the μ-slide chamber cell co-culture chemotaxis assay. HUVECs (indicated as ECs) were seeded in the central observation area whereas mock- and ACKR3-CHO cells are seeded in the left and right lateral reservoirs, respectively. When CXCL12 is added to both reservoirs (left panel), ACKR3- CHO cells act as chemokine scavenger and generate a chemotactic CXCL12 gradient for HUVECs. No migration was observed when CXCL12 was replaced by vehicle (right panel). (B,C) Leftward (black symbols) and rightward (red symbols) trajectory plots of the migration of individual HUVECs exposed to the experimental conditions described in (A). Blue dot: centre of mass. (D) Quantification of the corresponding forward migration index (FMI) and of the displacement of centre of mass (COM) parameters. Data represent the mean ± S.E.M. of three independent experiments. ^**p < 0.01, Student t-test.