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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: J Immunol. 2019 Mar 27;202(9):2585–2608. doi: 10.4049/jimmunol.1801350

Fig. 5. A pro-inflammatory macrophage polarization but not a neutrophilic influx at the maternal-fetal interface and myometrium prior to in vivo T-cell activation-induced preterm birth.

Fig. 5.

(A) Gating strategy used to identify M1-like (CD11b+F4/80+iNOS+ cells) macrophages at the maternal-fetal interface. Grey histograms represent autofluorescence controls and colored histograms represent the expression of iNOS at the maternal-fetal interface of dams injected with isotype control or αCD3ε, respectively. Proportions of M1-like macrophages in the decidual and myometrial tissues from dams injected with αCD3ε or isotype, (n=12-13 each). (B) Left to right: spatial localization of the murine decidua and myometrium. Workflow showing the magnetic isolation of macrophages from decidual and myometrial cells; macrophage purity (F4/80+ cells; >92%) was determined by flow cytometry. (C) Heat map visualization of the expression of M1 macrophage markers by F4/80+ cells isolated from the decidual and myometrial tissues of dams injected with αCD3ε, isotype, LPS, PBS, RU486 or DMSO (n=6-8 each). (D) Heat map visualization of the expression of M2 macrophage markers by F4/80+ cells isolated from the decidual and myometrial tissues of dams injected with αCD3ε, isotype, LPS, PBS, RU486 or DMSO (n=6-8 each). Negative (−)∆Ct values were calculated using Actb, Gusb, Gapdh and Hsp90ab1 as reference genes. (E) Gating strategy used to identify neutrophils (CD45+Ly6G+ cells) at the maternal-fetal interface. Proportion of neutrophils in decidual and myometrial tissues from dams injected with αCD3ε, isotype, LPS, PBS, RU486 or DMSO (n=9-13 each). The p-values were determined by 2-tailed Mann-Whitney U-test. Data are shown as scatter plots (median).