Fig. 5

EV-IL-32 promotes macrophage M2 polarization. A Schematic chart showed MDMs cocultured with EV treated with or without GW4869. B MDMs cocultured with EV (50 µg/mL) isolated from EC109 shNC, EC109 shIL-32, KYSE150 vector and KYSE150 IL-32β for 72 h. The percentage of CD14⁺CD206⁺ macrophages were tested by flow cytometry. C The EV were derived from EC109 shNC cells which were treated with or without GW4869 at 10 µM. Western blot analyzed the expression of TSG101 in EV. D EV-IL-32 in lysate was detected by ELISA. E MDMs cocultured with EV (Derived from 1 × 10⁷ EC109 shNC cells treated with or without GW4869) for 72 h. Flow cytometry analysis of the CD14⁺CD206⁺ macrophages. F The EV derived from KYSE150 IL-32β cells treated with or without GW4869 at 10 µM. Western blot analysis of the expression of TSG101 in EV. G EV-IL-32 in lysate was detected by ELISA. H MDMs cocultured with EV (Derived from 1 × 10⁷ KYSE150 IL-32β cells treated with or without GW4869) for 72 h. Flow cytometry analysis of the CD14⁺CD206⁺ macrophages. I MDMs cocultured with EV (50 µg/mL) derived from EC109 shNC, EC109 shIL-32, KYSE150 vector and KYSE150 IL-32β for 48 h. The mRNA expression of M1 (IL-1β and iNOS) and M2 (IL-10 and Arg1) macrophage associated genes were detected by qRT-PCR. All data are representative of three independent experiments and represented as means ± SEM. Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001