Fig. 3

Both secretion and function of exosomal Rab22a-NeoF1 fusion protein are dependent on its binding to HSP90. a The co-IPs were performed using ZOS-M cells with anti-IgG, anti-mAb RAD5–8, or anti-HSP90 at their endogenous levels. WCL whole-cell lysate of ZOS-M cells. b The indicated stable 143B cells and ZOS-M cells were treated with the HSP90 inhibitor (Ganetespib) for 24 h, and then the exosomes were purified and analyzed by Western blotting. c The 293T cells were co-transfected with the indicated plasmids and then were lysed and analyzed by immunoprecipitation using S protein beads and Western blotting. d The Exosomes derived from the indicated stable 143B cells were isolated and analyzed by Western blotting. Data in a–d are representative of n = 3 biologically independent experiments. e The indicated cells were treated with exosomes derived from the indicated stable 143B cells for 24 h and then were subjected to migration and invasion assays. Data are mean ± s.d. of n = 3 biologically independent experiments. P values are shown. f–h Representative IVIS imaging (f), H&E-stained lung sections (g), and quantification of lung metastatic foci (h) from mice orthotopically injected 143B-Luc cells with exosomes derived from the indicated stable 143B cells. n = 5 biologically independent mice. Data are mean ± s.d. P values are shown. Scale bar, 2 mm