Fig 8.

17-DMAG reduces the EBV titer in lytically infected cells. AGS-Akata cells were transfected with SG5 or SG5-Z expression vectors, followed by a 48-h treatment with 17-DMAG (0.17 μM) or the DMSO control beginning at 4 h posttransfection. The cell medium was replaced after 48 h with medium free of drug. At 72 h posttransfection, cells and supernatant were harvested. (A) The virus titer was quantitated by infecting Raji cells with various amounts of the supernatant at 72 h posttransfection and counting the number of GFP-positive cells (green Raji units [GRU]) by using a fluorescence microscope. The results from two independent experiments are shown. (B) The number of viable AGS-Akata cells under each condition shown in panel A was determined by counting the cells (with trypan blue) at the end of the experiment. (C) Whole-cell extracts were prepared from the AGS-Akata cells used to derive the virus titer shown in panel A, and immunoblot analysis was performed to analyze the expression of BMRF1, BZLF1, and actin.