Fig 2: Downregulation of B7-H4 by T. gondii infection resulted in changes in dMf function by affecting the JAK2/STAT1 signaling pathways. a, b Representative western blot and histogram analysis of B7-H4, JAK2, p-JAK2, STAT1 and p-STAT1 levels of dMf in uninfected, infected and B7-H4 neutralized infected groups. c, d Representative western blot and histograms analysis of B7-H4, STAT1, p-STAT1, iNOS and TNF-a of dMf in the uninfected, infected, infected + STAT1 inhibitor (Infected + Flu), B7-H4 neutralized infected and B7-H4 neutralized infected + STAT1 inhibitor (Neutralized infected + Flu) groups. The data in all panels are representative of at least three independent experiments. Data are presented as the means ± SD, with the unpaired t-test. Asterisks indicate significant differences between groups at *P < 0.05, **P < 0.01. Flu, Fludarabine; JAK2, Janus kinase 2; p-JAK2, phosphorylated JAK2; p-STAT1, phosphorylated signal transducer and activator of transcription 1; STAT1, signal transducer and activator of transcription 1

Fig 3: Reduction of B7-H4 expression on dMf by T. gondii infection affects the expression of M1- and M2-type membrane functional molecules. a-e Flow cytometry analysis of CD80, CD86, CD206 and CD163 levels on human dMf in uninfected, infected and B7-H4 neutralized infected groups. f-i Flow cytometry analysis of B7-H4, CD80, CD86 and CD206 expressions on dMf in uninfected, infected, and B7-H4-/- infected mice. Data are presented as the mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by unpaired the t-test. Asterisks indicate significant differences between groups at *P < 0.05, **P < 0.01. PE, Phycoerythrin; SSC, side scatter

Fig 4: Reduction of B7-H4 expression on dMf with T. gondii infection resulted in changes in the expression of TNF-a and IL-10. a-c Expression level of IL-10 and TNF-a in F4/80+ dMf in uninfected, infected and B7-H4-/- infected mice detected by flow cytometry. d, e Western blotting assay of TNF-a and IL-10 levels in CD14+ dMf of the uninfected, infected and B7-H4 neutralized infected groups. f, g Representative immunofluorescent photographs of B7-H4, TNF-a and IL-10 of CD14+ dMf in the uninfected, infected and B7-H4 neutralized infected groups. Scale bar: 50 µm. Data are presented as the mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by the unpaired t-test. Asterisks indicate significant differences between groups at *P < 0.05, **P < 0.01. DAPI, 4',6-Diamidino-2-phenylindole; IL, interleukin; TNF, tumor necrosis factor

Fig 5: Changes in B7-H4 expression on decidual macrophages (dMf) and the effect of these changes on phagocytic activities during Toxoplasma gondii infection. a B7-H4 expression levels on human CD14+ dMf in uninfected, infected and B7-H4 neutralized infected groups detected by flow cytometry. b B7-H4 expression on mouse F4/80+ dMf detected in uninfected, infected and B7-H4-/- infected mice by flow cytometry. c Fluorescence microscopy images of the number of phagocytic dMf in the uninfected, infected and B7-H4 neutralized infected groups. Scale bar: 50 µm. d Flow cytometry results showing the phagocytotic capacity of dMf in the uninfected, infected and B7-H4 neutralized infected groups. Data are presented as mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by the unpaired t-test. Asterisks indicated significant differences between groups at *P < 0.05, **P < 0.01. IgG-FITC, immunoglobulin G-fluorescein isothiocyanate complex