M1-Mφ suppress EMT, migration, and invasion of trophoblasts via secreting EVs. (A) Schematic illustration of the EV acquisition method and the experimental design. (B) Representative TEM image of M1-EVs with a lipid bilayer structure (Scale bar, 100 nm). (C) NTA of the size distribution and concentration of M1-EVs. (D) Western blot analysis of EV fractions and cell lysates of M1-Mφ with antibodies against exosomal proteins (CD63, CD81, TSG101) and the cellular protein calnexin. (E-F) Western blot and RT-PCR analysis of E-cadherin (E-cad), N-cadherin (N-cad), and vimentin (Vim) protein and mRNA levels in HTR-8 and JEG3 cells treated with M1-EVs or an equal volume of medium from M1-Mφ treated with GW4869. (G-J) Cell migration and invasion capacities of trophoblasts (HTR-8 and JEG3) treated with control, M1-EVs, or EVs from an equal volume of medium from M1-Mφ treated with GW4869 determined by wound healing and transwell assays, respectively. Representative images of migrated or invaded cells are shown (magnification, × 200). Error bars, SD. *P < 0.05, **P < 0.01, ***P < 0.001.