Fig. 6

EVs from cervical cancer patients after radiotherapy contribute to increased macrophage phagocytosis. Peripheral blood mononuclear cells from healthy donors were obtained and cultured in RPMI 1640 medium. M2-polarized macrophages were obtained by IL-4+IL-13 stimulation and treated with EVs from cervical cancer patients before and after radiotherapy. A, representative scatter diagrams of flow cytometry analysis the expression of PD-1 and CCR7 in macrophages (CD45⁺CD14⁺CD11b⁺) were shown. Histograms of PD-1⁺ cells in total macrophages (B) and in CCR7⁺ macrophages (C) were shown. Macrophage phagocytosis was assessed using pHrodo Red E. coli bioparticles or CFSE labeled EVs. Representative diagram (D) and histogram (E) about mean fluorescent intensity (MFI) of pHrodo in macrophages were shown. Representative scatter diagram (F) and histogram (G) of flow cytometry analysis the correlation between PD-1 expression and MFI of pHrodo in macrophages were shown. Representative diagram (H) and histogram (I) of CFSE MFI in macrophages co-cultured with EVs labeled with or without (Control) CFSE, after treated with EVs from cervical cancer patients before and after radiotherapy for 24 hours, were shown. Data were presented as mean±SD. P value was calculated by two-tailed Mann Whitney U test