Figure 3.

LPA promotes the adhesion and enrichment of M2-polarized macrophages in decidua. (a) Heatmap of differential genes in RNA-seq for pMo (n = 3) and dMφ (n = 3) from women with normal pregnancy. (b) Enriched KEGG and GOTERM pathways of differential gene clustering from (a). (c and d) Adhesion assays for control or LPA-treated (24 h) PKH-26-labeled red-fluorescent M0 macrophages to PKH-67-labeled green-fluorescent DSCs (n = 5). (e and f) Adhesion assays for PKH-26-labeled red-fluorescent M2 macrophages treated with vehicle (n = 10), ENPP2 inhibitor (n = 7), LPAR1 inhibitor (n = 6) or PPARG inhibitor (n = 6) for 24 h to PKH-67-labeled green-fluorescent DSCs. (g and h) The residence and number of ADGRE1⁺ macrophages in the uterus of WT (n = 5) and Enpp2^± (n = 4) mice by immunofluorescence assays. (i and j) Flow cytometry assays for detection of the proportion in PTPRC⁺ immune cells and the absolute numbers of ADGRE1⁺ITGAM⁺ uterine macrophages per 10,000 cells in uterus of WT (n = 8) or Enpp2^± (n = 8) pregnant mice. (k and l) Adhesion molecules and differentiation markers of PTPRC⁺ADGRE1⁺ITGAM⁺ uterine macrophages of WT (n = 8) or Enpp2^± (n = 8) pregnant mice in flow cytometry assays. Data were presented as mean ± SEM or median and quartile and analyzed by t test, Mann-Whitney U test or one-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001.