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. 2022 Apr 19;3(2):101337. doi: 10.1016/j.xpro.2022.101337

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© 2022.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Gating strategy for FACS-sorting of wound macrophages

(A and B) Representative flow cytometry plots at (A) 4 and (B) 14 days post injury. DAPI^- single cells were gated (Gates G1-G4) and expression of CD45 was analyzed. CD45⁺ cells were gated (G5) and analyzed for expression of CD11b and F4/80. Single CD45⁺CD11b⁺F4/80⁺ wound macrophages (G6) were sorted in 384-well plates for RNA sequencing. CD45⁺CD11b⁺F4/80⁺ wound macrophages can further be analyzed for expression of Trem2, CD301b, and MHC II. Index sorting allows merging FACS data with gene expression data at single cell level.