Fig. 2: The amounts of TβRII⁺ crEVs positively correlate with tumor burden.

a Experimental analysis in vivo: mice were not inoculated (−) or were given subcutaneous inoculation (+) of MCF10A-RAS cells (5 × 10⁵ cells per mouse) expressing control shRNA or doxycycline-inducible shRNA targeting TβRII (shTβRII) and tumors were grown for 3 weeks, followed by the administration of doxycycline (Dox). 3 d later, mice were euthanized for further analysis (n = 10 mice per group). b Schematic of ELISA of human TβRII on EVs in plasma samples from mice with human breast cancer xenograft. c FACS analysis (left) of the percentage of TβRII⁺ crEVs and ELISA (right) of TβRII on crEVs in plasma samples from mice (n = 10 mice per group). d Pearson correlation between the TβRII⁺ crEVs or TGF-β1 in plasma and the tumor burden from the control group mice without Dox administration (n = 10 mice per group). e Quantification of circulating EV-TβRII at the indicated tumor volume (left panel). Pearson correlation between the circulating EV-TβRII in plasma and tumor burden from mice was shown right (n = 5 mice per group). f Schematic diagram of longitudinal blood collection pre-operatively, post-operatively or relapse. g Electron microscopy images of EVs secreted from healthy donors, patients with pre- and post-operatively breast cancer. Scale bars, 50 nm. h FACS (left) and ELISA (right) of TβRII⁺ crEVs in plasma samples from patients with breast cancer pre-operation (n = 6 patients), post-operation (n = 6 patients), relapse (n = 3 patients) and healthy donors (n = 20 samples). *p < 0.05 (two-tailed Student’s t test (c, e (left), h) or two-way ANOVA (d, e (right))). Data are analyzed of three independent experiments and shown as mean ± SD (c, e (left), h). Source data are provided as a Source Data file.