Figure 3

XMRV induced downregulation of A3G in prostate cancer cells We used culture supernatant from chronically infected LNCaP cells with XMRV as the source of infectious XMRV. Virus infections were performed using cells plated 1 day before infection. Cells were at 50% confluency at the time of infection. On the day of infection, fresh media containing 5 μg/ml polybrene was added to the cells and virus was layered on the cells and incubated for 6 h to allow virus adsorption. Cells were then washed once with PBS, and fresh media containing FBS were added. After 1-2 weeks, XMRV infection was confirmed by detecting XMRV p30 protein by using goat polyclonal anti-Rauscher MLV p30 Gag in uninfected (-) and XMRV infected (+) LNCaP cells (A) and DU-145 cells (B). (C) A3G expression was determined by the protocol described in Figure 1 using anti-ApoC29. (D) Densitometry of A3G expression as determined by three independent experiments.