< Back to Article

ESCRT-Independent Budding of HIV-1 Gag Virus-Like Particles from Saccharomyces cerevisiae Spheroplasts

Figure 2

Characterization of VLPs.

(A) VLPs were collected from wild type cells expressing wild type Gag-GFP and subjected to centrifugation across a sucrose gradient (20–70%, weight to volume). 1 ml fractions were subsequently collected, the density of each fraction was measured, and each fraction was subjected to SDS-PAGE and anti-GFP immunoblotting. (B) Wild type cells expressing either Gag-GFP (WT) or (G2A) were equivalently processed to isolate the whole cell and VLP fractions. The VLP fraction was subsequently incubated with either PBS alone (PBS), PBS+Trypsin (T), or PBS+Trypsin+Triton X-100 (T+TX100), and post reaction material was visualized by immunoblotting using anti-GFP antibody. Whole cell lysates (W.C.) from cells expressing either WT Gag-GFP or used to generate the VLP fraction were loaded onto the gel to indicate the relative amount of expression.

doi: https://doi.org/10.1371/journal.pone.0052603.g002