Figure 5. Biological effects of secreted Cav-1.

(A) Western blot analysis of secreted Cav-1 in the concentrated conditioned medium from some representative melanoma cell lines (left) and Cav-1-transduced WM983A (right). (B) Effects of the conditioned medium collected from the WM983A/LXSN (Control medium) or from the WM983A/Cav-1 (Cav-1CM) on the invasion and migration abilities of WM983A/LXSN cells. The polyclonal anti-Cav-1 antibody was added to the conditioned media, where indicated, at increasing doses from 2 to 10 μg/ml, *p≤0.05; ** p≤0.001 vs control - °p≤0.05; °°p≤0.001 vs Cav-1 CM; (C) Invasion and migration assays in Me1811 melanoma cell line performed with increasing doses of Cav-1 antibody added in the cell medium (concentrations of the anti-Cav-1 Ab were 4, 10, 20 and 40 μg/ml). All the results are reported as mean±SD. Each experiment was performed in triplicate. An anti-IgG Ab was included as a control. (D) (left and middle) Western blot analyses on microvesicles purified from the indicated melanomas; Ponceau S indicates the equal protein loading. (right) Normalized amounts of microvesicles released from the same samples. (E) Invasion and migration assays in control- or Me665/1 MV-fused WM983A cells; when indicated 10 μg/ml of an anti-Cav-1 Ab was included. (mean±SD derives from at least 3 independent experiments), ** p≤0.001 vs WM983A.