Western blot analysis of Akt phosphorylation in different EBV-infected cell lines.
(A) Analysis of activated Akt phosphorylation at Ser473 and Thr308 in ER-EB2.5 cells in the absence of β-estradiol (−) or 17 h after β-estradiol re-addition (+). Blots were also probed with a pan Akt antibody to detect total Akt levels and anti-LMP1 antibodies. Actin was used as a loading control. Numbers under panels show quantification of these representative western blots with signals for each antibody normalised to actin and expressed relative to ER-EB 2.5 (−β-estradiol). (B) Western blot analysis in the BL31 cell line series infected with either wild-type recombinant EBV (BL31 wtBAC3, BL31 wtBAC2), EBNA3A knock-out EBV (BL31 3AKO-3AΔ, BL31-3AKO-1.1), EBNA3B knock-out EBV (BL31-3BKO-1, BL31-3BKO-8.2), EBNA3C knock-out EBV (BL31-3CKO-6, BL31-3CKO-3), EBNA3 knock-out EBV (BL31-E3KO) or with their respective revertant viruses (BL31-3A rev-2, BL31-3B rev-2.2, BL31-3C rev-2 and BL31-E3 rev). Non-specific bands (*). (C) Western blot analysis of additional BL31 knock-out cell lines (D) Western blot analysis of a conditional LCL expressing an EBNA3C-HT fusion protein cultured in the absence of HT for 21 days and then with HT re-added for 10 days. Numbers under panels show quantification of these representative western blots with signals for each antibody normalised to actin and expressed relative to −HT.