LMP1 provides a proliferative advantage that can be eradicated following PARP inhibition.

A) IPA analysis predicts LMP1 to activate proliferation pathways and PARP inhibition to inactivate proliferation pathways. B and C) Cells were stained by CFSE (5(6)-Carboxyfluorescein N-hydroxysuccinimidyl ester) and allowed to proliferate for 96 hrs- LMP1+ vs LMP1- CFSE labeled cells and LMP1+ untreated cells vs olaprib-treated LMP+ CFSE labeled cells were then detected by FACS analysis, respectively. D) Cell cycle analysis- LMP1+ cells were incubated with 1 μM olaparib for 72 hrs. Cells were then harvested, fixed and permeabilized in absolute ethanol and then incubated with propidium iodide (PI) and RNAse A for 30 mins at 37C and analyzed by FACS. E and F) Methylcellulose colony forming cell (CFC) assay- 500 LMP1+ cells, untreated and pre-treated with 2.5 μM olaparib for 96 hrs, were seeded in methylcellulose media and incubated for 14 days at 37°C. Images were captured following staining with crystal violet and unstained at 4X magnification, respectively.