Fig. 4.

The functions of exosomes containing Rab22a-NeoF1 fusion protein on its negative recipient cancer cells requires its binding partner PYK2 from donor cells. a, b The exosomes derived from the indicated stable 143B and U2OS (a), as well as ZOS-M (b) cells were purified and subjected to Western blotting. c The co-IPs were performed using ZOS-M cells with anti-mAb RAD5–8, anti-PYK2, or anti-IgG at their endogenous levels. WCL whole-cell lysate of ZOS-M cells. d The 143B cells were treated with the indicated exosomes for 1 h and then were lysed. Rho-GTP forms were pulldown by RBD beads and were subjected to Western blotting. The relative intensity of RhoA-GTP was quantified with the ImageJ software and normalized with RhoA-Total. Data in a–d are representative of n = 3 biologically independent experiments. e The 143B cells were treated with the indicated exosomes 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 the indicated exosomes. n = 5 biologically independent mice. Data are mean ± s.d. P values are shown. Scale bar, 2 mm