HDAC inhibitor–mediated induction of EBV lytic-phase gene expression. (A) Comparison of TK and BGLF4 transcript expression in P3HR1 cells after exposure to different HDAC inhibitors. Cells were treated with individual HDAC inhibitors as indicated and total RNA was analyzed by reverse transcription and real-time PCR analysis as described in the legend to Figure 2. Expression of β-actin mRNA under similar treatment conditions was used to normalize the dataset. (B) Immunoblot analysis for EA-D protein. Thirty micrograms of whole-cell lysates from individual HDAC inhibitor–treated cells (for 48 hours) or untreated cells were separated in 10% SDS-PAGE, transferred to a nitrocellulose membrane, and immunoblotted with a 1:2000 dilution of mouse anti-EBV EA-D Ab (Millipore). Equal loading of proteins was verified by immunoblotting with 1:15 000 dilution of mouse anti–β-actin Ab (Sigma-Aldrich). Each assay was repeated 3 times and error bars in each individual figure represent SDs.