Hsp72 association and coactivation with EBNALP or EBNALP C-terminal deletion mutants in BJAB cells. (A) Schematic depiction of N-terminally Flag-tagged EBNALP or EBNALP C-terminally deleted mutant proteins, FLP, FLPd10, FLPd34, and FLPd45. (B) EBNALP associates with Hsp72 and EBNA2 does not. BJAB cells were transfected with 10 μg FHsp72 expression vector and with 10 μg EBNA2 or EBNALP expression vector. Lysates were immune precipitated with PE2 (E2) or JF186 (LP) followed by Western blot analysis with M2, PE2, or JF186 antibody. Five percent of FHsp72 input is shown. (C) Hsp72 associates with FLP or FLPd10, but not with FLPd34 or FLPd45. BJAB cells were transfected with 10 μg FHsp72 expression vector or empty expression vector and with 10 μg FLP, FLPd10, FLPd34, FLPd45, or control expression vector. Lysates were immune precipitated with JF186 antibody to EBNALP followed by immune blot analysis with M2 or JF186 antibody. Five percent of FHsp72 input is shown. (D) Hsp72 augments EBNALP, but not EBNALPd45, coactivation with EBNA2. BJAB cells were transfected with 10 μg EBNA2, EBNALP, or EBNALPd45, and FHsp72 expression vector, 5 μg LMP1p-Luciferase Reporter, 2 μg β-gal control plasmid, and sufficient pSG5 vector to achieve equivalence in DNA input. Assays were done after 24 hours.