Skip to main content
. 2003 Sep;77(17):9542–9552. doi: 10.1128/JVI.77.17.9542-9552.2003

FIG. 6.

FIG. 6.

Association of HSV receptors with detergent-insoluble low-density sucrose fractions containing rafts. 3E5 cells stably expressing HVEM-GFP (A), C10 cells stably expressing nectin-1 (B), and 293T cells transiently transfected with a plasmid expressing the HVEM (C) or nectin-1 (D) gene were homogenized in Triton X-100 and fractionated on sucrose gradients. Proteins (10 μl of each fraction) were resolved on SDS-PAGE and transferred to nitrocellulose, and the distribution of HSV receptors was analyzed by Western blotting with MAb CW10 to HVEM or MAb CK6 to nectin-1. Fraction numbers are indicated at the top of the figure (P for pellet). T and B refer to the top and bottom of the gradient, respectively. The distribution of the known raft-associated protein flotillin-2 (flot-2) was analyzed with a commercial MAb and served as a positive control for rafts (A, B, C, and D, middle panels). Also shown is the distribution of the raft-specific ganglioside GM1 analyzed by dot blot with CTB-HRP (A, B, C, and D, lower panels). Sizes are shown on the left (in kilodaltons).