Figure 2

Z-eGFP virus like particles detection and characterization. A. Western Blot with anti-Z inmunoglobulines of Z-eGFP expression in 293T cells. Lane 1. 293T untransfected monolayer (control). 2. 293T monolayer transfected with pZ-eGFP. (Cell) Cell lysate 3. Pellet of the ultracentrifugated supernatant of untransfected 293T cells (control). 4. Pellet of the ultracentrifugated supernatant of 293T cells transfected with pZ-eGFP. (Sup) pelleted material from supernatant. B. Protease protection assay of Z-protein-containing particles in the supernatant of pZ-eGFP transfected 293T cells. Western Blot with anti-Z inmunoglobuline of the pellet of the ultracentrifugated supernatant of transfected cells, treated with proteinase K to final concentration of 0.04 μg/μl, 0.08 μg/μl and 0.4 μg/μl, with or without Triton X-100, or untreated (lane 1). C and D. Transmission electron microscopy with immunogold labelling of the pellet obtained by ultracentrifugation through a sucrose cushion of the supernatant of pZ-eGFP transfected cells. The red arrow indicates the gold labelling. The bar in each figure represents 100 nm.