Fig 2: Dynamical changes of monocyte subsets after myocardial infarction. (A) Representative illustration of three monocyte subsets according to CD14 and CD16 staining at different time-points. (B) Quantitative analysis indicated that the total cell counts initially increased (3–5 h) and then declined almost to the level of healthy controls (left panel in B). The right panel in B displays the proportional changes of three subsets of monocytes over time, revealing the progressive increase of the percentage of the IM subpopulation. *P<0.05, total cell counts vs. healthy; **P<0.01, proportion of IM vs. healthy. CM, classical monocytes; NC, non-classical subsets; IM, intermediate subpopulation; PE, phycoerythrin; APC, allophycocyanine; Cy, cyanine; FSC, forward scatter; SSC, side scatter; Q, quadrant.

Fig 3: Patients RM exhibit weakened interactions between DSCs and CD16+ dMφ. (A) Compared the expression of COL4A1 in decidua from women with normal pregnancy (NP, n=8) or recurrent miscarriage (RM, n=8) by Immunohistochemistry. Scale bar, 50 μm. (B) Compared the expression of OPN in decidua from women with NP (n=10) or RM (n=10) by Immunohistochemistry. Scale bar, 50 μm. (C) Compared the expression of HA in decidua from women with NP (n=8) or RM (n=8) by Alcian blue staining assays. Scale bar, 50 μm. (D) Levels of CD16, CD86, CD206 and CD209 in CD45+CD14+ dMφ from women with NP (n=7) or RM (n=7) by flow cytometry assays. The data are presented as the mean ± SEM; two-tailed Student’s t-test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig 4: CD16+ macrophages from endometrium and decidua exhibit higher levels of adhesion molecules. (A) Expression of CD49a, CD44 and LYVE1 in CD45+CD14+CD16+ or CD45+CD14+CD16- eMφ (n=12) in flow cytometry assays. (B) Expression of CD49a, CD44, LYVE1 and Integrin αVβ3 in CD14+CD16+ or CD14+CD16- dMφ (n=11) in flow cytometry assays. The data are presented as the mean ± SEM; two-tailed Student’s t-test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns.

Fig 5: Exogenous COL4A1, OPN and HA promote development of immunomodulatory CD16+ Mφ. (A, B) After 48 hours of stimulation with exogenous COL4A1 (10 ug/mL), OPN (1 ug/mL) or vehicles, the expression levels of CD16 on U937-induced Mφ (A; n=5) and the polarization-related molecules CD86, CD209 and CD206 on CD16- or CD16+ Mφ (B; n=5) were detected by flow cytometry. (C, D) After 48 hours of stimulation with exogenous HA (0, 50, 100 μM), the expression levels of CD16 on U937-induced Mφ (C; n=4) and CD86, CD209 and CD206 on CD16- or CD16+ Mφ (D; n=4) were detected by flow cytometry. The data are presented as the mean ± SEM; oneway ANOVA test or two-tailed Student’s t-test; *P < 0.05, **P < 0.01, ****P< 0.0001, NS, no significant difference.