Figure 1

Macrophages from Ltb4r2−/− mice exhibit reduced chemotaxis. Bone-marrow-derived macrophages (BMDMs) were isolated from WT and Ltb4r2−/− mice and unpolarized (M0) or polarized to M2-like and M1-like states. A, schematic representation of experimental design; (B) iNOS+ cells as a percentage of F4/80+ CD11b+ cells as determined by flow cytometry; (C) CD206+ cells as a percentage of F4/80+ CD11b+ cells as determined by flow cytometry; (D) principal component analysis plot of RNA sequencing data; (E) Volcano plot showing differentially expressed genes by p value in M0 macrophages (each dot represents a distinct gene); (F) Volcano plot showing differentially expressed genes by p value in M2-polarized macrophages; (G) Volcano plot of differentially expressed genes by p value in M1-polarized macrophages; (H) Enrichr Gene Ontology pathway analysis of differentially expressed genes in M1-polarized macrophages, ordered by p-value; (I) relative expression of Ccl5 and Lgals3 in M1-like macrophages from WT and Ltb4r2−/− mice; (J) representative images of macrophage migration in transwell plates (left panels) and respective quantification (right panel). Arrows indicate macrophages (blue) in left panel images. Scale bar = 500 μm; (K) quantification of migrated macrophages after treatment with vehicle (V) or 6 μg LY255283 (LY). All data are presented as mean ± SEM and each data point represents an independent biologic replicate from different animals. For statistics, unpaired two-tailed t tests were used for comparison of two values, and 1-way ANOVA with Tukey post-test for more than two comparisons.