Fig. 5
The functions of exosomal Rab22a-NeoF1 fusion protein requires recruiting BMDMs to the pulmonary pre-metastatic niche. a Schematic illustration for pre-educated tumor-free mice model. b The indicated gene mRNA levels from the lungs of mice treated with the indicated conditioned media (CM) were quantified by qPCR. n = 5 biologically independent mice. Data are mean ± s.d. P values are shown. c Mice lungs pre-educated with the indicated conditioned media (CM) were homogenized and analyzed by flow cytometry for numbers of both CD11b+ cells and F4/80+ cells. n = 5 biologically independent mice. Data are mean ± s.d. P values are shown. d Representative immunofluorescent analyses of CD11b and F4/80 expression in the lungs from mice treated with the indicated conditioned media (CM). Scale bar, 100 μm. e–g Mice were pre-educated with the indicated conditioned media (CM) for 3 weeks, and then were orthotopically or tail vein injected with 143B-Luc cells for the indicated periods. The lung metastases were analyzed. Representative IVIS imaging (e), H&E-stained lung sections (f), and quantification of lung metastatic foci (g). n = 4 biologically independent mice. Data are mean ± s.d. P values are shown. Scale bar, 2 mm. h–j Mice orthotopically co-injected U2OS/MTX300-Luc cells with either ZOS-M-shNC cells or ZOS-M-shRAB22A-NeoF1 cells at the 10:1 ratio, and then intravenously injected liposome or PBS twice a week for 3 weeks and the lung metastases were analyzed. Representative IVIS imaging (h), H&E-stained lung sections (i), and quantification of lung metastatic foci (j). n = 5 biologically independent mice. Data are mean ± s.d. P values are shown. Scale bar, 2 mm
