Fig. 3

EV-dead promotes the invasion and BCSC subpopulation of the co-cultured human TNBC MDA-MB-231 cells by inducing macrophage M2 polarization. A Diagram of the EV-dead^ADR and EV-alive-H separation procedures and their representative TEM images. Scale bar: 100 nm. B EV protein markers were identified by Western blotting; (C) EV sizes and concentrations were detected by NTA analysis; (D) CXCL1 concentrations in EV-dead^ADR and EV-alive-H were detected by an ELISA assay; (E, F) The polarization changes of THP1 macrophages after EV-dead^ADR treatment (25–100 μg/ml) for 48 h; 50 μg/ml EV-dead was used for the Western blotting assay; G Invasion efficacy changes of the co-cultured MDA-MB-231 cells after EV-dead^ADR treatment as indicated. Scale bars: 100 μm; H Changes in apoptosis resistance of the co-cultured MDA-MB-231 cells to paclitaxel after EV-dead^ADR treatment for 48 h, as determined by annexin V-FITC/PI staining and flow cytometry; I The ALDH⁺ BCSC subpopulation and CD133⁺ BCSC subpopulation in the co-cultured MDA-MB-231 cells after EV-dead^ADR treatment for 48 h were detected by flow cytometry; n = 3. Data are presented as mean ± SD. ^*p < 0.05, ^**p < 0.01