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. 2008 May 16;4(5):e1000065. doi: 10.1371/journal.ppat.1000065

Figure 2. PRV Us9 and gB are highly-enriched in DRMs of non-polarized PC12 cells, whereas gH is not.

Figure 2

Non-differentiated PC12 Cells were infected with PRV Becker for 12 hours, and then lysed with cold 1% Triton X-100. Lysates were separated on a discontinuous Optiprep™density gradient, and 10 fractions were collected from the top to the bottom of the tube (1 ml each). Samples 2–9 were subjected to SDS-PAGE, and Western blotting analysis was performed using biotinylated cholera toxin B subunit (for GM1) and antibodies to PRV Us9, gB, gE, gC, gH and transferrin receptor (TfR). To test the effect of cholesterol depletion on Us9 association with DRMs, infected PC12 cells were incubated with 20 mM methyl-cyclodextrin (MCD) at 37°C for 45 minutes prior to lysis with cold detergent. The preprocessed form of gB (*) is labeled along with the 69 kDa and 58 kDa (−) subunits.