Figure 4.

Glypican-3 in human serum small EVs. (a) Schematic representation of sEVs extraction. Pellets were used for size and morphology characterization using Nano Sight NS3000 and TEM, respectively. Protein contents were evaluated with western blot. (b) Representative TEM image of sEVs isolated from HCC serum (scale bar: 0.1 µm). (c) GPC-3 sEVs content was evaluated by WB in healthy controls and HCC cases. GPC-3_C (red-top panel) and GPC-3_N (green-bottom panel) antibodies were used for immunoblotting. (d) Upper blot—Merged immunoblot of N- and C-terminal GPC-3 IBs, Lower Blot—GPC-3_N IB is represented as well in black/white. (e) schematic representation for serum sEVs and supernatant isolation to verify GPC-3 soluble content. (f) IB of GPC-3_N in sEVs and corresponding supernatant (Sup) in three HCC cases and 1 control.