NETs promote fibroblast differentiation. (A) Characterization of differentiated cells after coincubation of CTEPH monocytes with dsDNA, isolated NETs, NETs plus DNase, or vehicle for their surface expression of TLR4, CD163, CD206, and fibroblast-specific protein (FSP) by flow cytometry (n = 4). The percentage of positive cells of single events was computed. (B) Representative coimmunofluorescent staining of α smooth muscle actin (α-SMA) (red) and fibroblast activation protein (FAP) (yellow) in CTEPH monocytes exposed to NETs (scale bars, 100 µm). (C) qPCR analysis for TGF-β signaling pathway genes in differentiated cells. Data are shown as mean ± SEM. *P < .05 (1-way ANOVA with Tukey’s post hoc correction).