Figure 2

Inhibition of mTORC1 alters early lytic replication in EBV-positive cells. A. AGS-BDneo or Raji cells were treated with 0, 1, 5, or 10 nM rapamycin 24 hr prior to induction of lytic replication. Western blot analysis was performed with anti-BMRF1, anti-Z, anti-R, and anti-tubulin antibodies. Representative blots are shown. B. The BMRF1 levels from Western blots (in triplicate) were quantified and standardized to tubulin levels. The resulting BMRF1 protein levels are presented, relative to induced cells (with no rapamycin treatment). C. BMRF1 levels of a panel of EBV-positive cells treated with 5 nM rapamycin, induced, and analyzed as in parts A and B above. Dark bars are epithelial cell lines, light bars are B cell lines. D. Z and R levels from the Western blots in part A were quantified and standardized to tubulin levels. The resulting Z and R protein levels are presented, relative to induced cells (with no rapamycin treatment). E. Inhibition of mTOR with 5 nM rapamycin inhibits mTOR activity. Raji and AGS-BDneo cells were treated with 0 or 5 nM rapamycin for 2 days, then subjected to Western blot analysis with anti-phospho-p70S6K and total p70S6K antibodies.