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. 2004 Apr 22;23(10):2175–2184. doi: 10.1038/sj.emboj.7600219

Figure 3.

Figure 3

Kinetics of release and oligomeric structure of soluble GP. (A) RK-13 cells infected with vSCGP8 were pulse-chase labeled and proteins were immunoprecipitated from cells (left panel) and medium (right panel). (B) Surface biotinylation of RK-13 cells expressing EBOV GP. Release of biotin-labeled GP from the cell surface into culture supernatant was observed at indicated time intervals. (C) Analysis of the oligomeric structure of shed GP. Vero E6 cells were infected with EBOV. At 5 days p.i., virus from culture medium was pelleted through a 20% sucrose cushion, and subsequently the supernatant and virus were analyzed by ultracentrifugation at linear sucrose gradients in the presence of 1% NP-40 and 0.4% DOC. Fractions 1–15 were collected from the bottom and analyzed by Western blot. Due to the effect of several detergents present in the samples such as NP-40, DOC, and finally SDS, the bands appeared slightly more diffused compared to other analyses.