Figure 3.

PP13 polarizes neutrophils toward a regulatory phenotype. (A) Expression of CD66 or CD11b on the surface of neutrophils co-cultured with BeWo cells and exposed to PP13 (3 μg/ml) (n = 3) or left untreated (n = 3) for 24 h (red: control, blue: treated with PP13). This is a representative flow cytometry experiment. (B) Migration assay of neutrophils in a transwell plate toward PP13, control Gal, or fMLP. (C) Expression of PD-L1 on the surface of neutrophils co-cultured with BeWo cells and exposed to PP13 (3 μg/ml) or left untreated for 24 h (n = 3) (red: control, blue: treated with PP13). This is a representative flow cytometry experiment. (D) De novo synthesis of TNFA mRNA in neutrophils cultured with or without PP13 for 6 h (n = 6). Data are presented as ΔCt values (normalization to GAPDH). (E) ELISA assay for the secretion of HGF or VEGF-α by neutrophils treated with PP13 (3 μg/ml) or Control Gal for 6 h (n = 9). (F) ELISA assay for the secretion of MMP-9 or IL-4 by neutrophils treated with PP13 (3 μg/ml) for 6 h (n = 5) (^**p < 0.01). All analysis was performed with Student's t-test (number of groups are 2) or One-way ANOVA unpaired test (number of groups are more than 2), ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.