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. 2017 Nov 24;18(12):2521. doi: 10.3390/ijms18122521

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© 2017 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Leukocyte-gating strategy for the measurement of S1PRs in cells subpopulations. Leukocytes were identified in a SSC/FSC plot for granulocytes, lymphocytes, and monocytes selection (A); The lymphocyte gate was copied to a CD3/S1PR plot to identify T-cells expressing S1PRs (CD3⁺/S1PR⁺) (B); The granulocyte population was copied to a CD66b/S1PR plot identifying granulocytes that present S1PRs (CD66b⁺/S1PR⁺) (C); To identify monocytes subsets, monocytes were copied to a CD14/CD16 plot and divided into three populations: classical monocytes (CD14⁺⁺/CD16⁻), intermediate monocytes (CD14⁺⁺/CD16⁺), and non-classical monocytes (CD14⁺/CD16⁺⁺) (D); Each monocyte population were copied to a CD14/S1PR plot to evaluate the presence of S1PRs in these monocytes (E, F, and G, respectively).