Fig 1: PTX3 promotes M2-like macrophage polarization. A, B The mRNA expression of M2 and M1 markers in THP-1 macrophages treated with or without 50 ng/ml PTX3. C The mRNA expression of M2 markers in PTX3-treated THP-1 macrophages with 250 ng/ml IgG1κ or WHC-001 (αPTX3Ab). D The mRNA expression of Arg1, Cd206, and Il1b in bone marrow-derived macrophages treated with or without 100 ng/ml PTX3. E Flow cytometric analysis of M1- and M2-like macrophages in BMDMs co-cultured with MEFs-shVoid, -shPtx3#915,or shPtx3#916 for 48 h. The CD45 + CD11b + F4/80 + cells were characterized by M1-like (CD86 + CD206−) and M2-like (CD206 + CD86−) macrophages through flow cytometric analysis. F The mRNA expression of Ptx3, Arg1, and Cd206 in tamoxifen-induced Ptx3fl/fl or Ptx3fl/fl; Ubc-Cre bone marrow-derived macrophages (M-CSF-primed M0 BMDMs) following stimulation with 20 ng/ml mIL-13 and 20 ng/ml mIL-4 (M2 polarization). G The mRNA expression of IFNG and TNFA in activated Jurkat cells (stimulated with PMA + ionomycin) incubated with conditioned medium from untreated THP-1 macrophages (Ctrl-CM) or PTX3-treated THP-1 macrophages (PTX3-CM). H Angiogenesis of HUVECs incubated in serum-free medium (No cell-CM), conditioned-medium from untreated THP-1 macrophages (Ctrl-CM) or conditioned medium from PTX3-treated THP-1 macrophages (PTX3-CM). The relative angiogenic ability was determined by the number of branch sites/nodes of tubes per field of view. I Western blot analysis of recombinant wild-type and N220A mutant PTX3 proteins treated with or without PNGase. J The mRNA expression of M2 markers in THP-1 macrophages treated with or without 50 ng/ml recombinant N220A mutant PTX3 protein. p values were calculated by two-tailed paired Student’s t test, one-way ANOVA, or two-way ANOVA. ns represents nonsignificant difference,*represents a p value of < 0.05, **represents a p value of < 0.01, and ***represents a p value of < 0.001. Values on the plots are presented as the means ± SEMs. Data are combined from 3 to 5 independent experiments