FIGURE 7.

Lys-CoA-Tat inhibits TGF-β1-induced Smad3 acetylation and EBV-lytic cycle. A, Mutu-I, Kem-I, and Sav-I cells pretreated, respectively, with 10, 10, and 20 μm of Lys-CoA-Tat or 20 μm of Ac-DDDD-Tat for 1 h were stimulated with TGF-β1 (2 ng/ml) for 17 h. The cell lysates were immunoprecipitated (IP) with anti-Smad3 antibody, and acetylated Smad3 was detected by immunoblot with anti-acetyllysine antibody. Equal amounts of protein were separated by SDS-PAGE and analyzed by Western blotting with antibodies to ZEBRA, EA, VCA, phosphoSmad3, Smad3, phospho-Akt, Akt, and tubulin. B, viability assay was performed with the LIVE/DEAD reduced biohazard viability/cytotoxicity kit (Molecular Probes, Invitrogen). The ability of this kit to detect cell death was controlled using a treatment with 0.4 mm of MnCl₂ (not shown). C, 3 μg of total RNA from Mutu-I, Kem-I, and Sav-I cells pretreated, respectively, with 10, 10, and 20 μm of Lys-CoA-Tat or 20 μm of Ac-DDDD-Tat for 1 h and then stimulated with TGF-β1 (2 ng/ml) for 17 h, were reverse transcribed. cDNA coding for ZEBRA was then analyzed by PCR; cDNA of hypoxanthine-guanine phosphoribosyltransferase (HPRT) was used as an internal control.