Fig 7.

Effects of pharmacological inhibitors on BZLF1 expression. (A and B) Effects of pharmacological inhibitors on BZLF1 expression. Raji cells were treated with either vehicle (Control), 10 μM DZNep (DZ), 300 nM trichostatin A (TSA), 1 μM 5-aza-2′-deoxycytidine (5Az), or in combinations as indicated. As a positive control (T/A/B), Raji cells were treated with 20 ng/ml TPA, 1 μM A23187, and 5 mM sodium butyrate. For DZNep or 5-Aza-2′-deoxycytidine (5Az) treatment, cells were exposed to the reagent daily for 3 days. Treatment with other chemicals was conducted for 24 h. Real-time RT-PCR was carried out as described in Materials and Methods. Levels of BZLF1 (A) and EBNA2 (B) mRNAs were normalized to GAPDH mRNA levels and are shown as fold increase. Each bar represents the means and SD from three independent treatments. (C to G) Effects of pharmacological inhibitors on epigenetic modifications. (C and D) Raji cells treated with DZNep were subjected to ChIP assays using anti-H3K27me3 (C) and anti-H4K20me3 (D) antibodies, followed by DNA extraction and real-time PCR to detect DNA fragments using the indicated primers. (E to G) Raji cells treated with inhibitors in the same way were subjected to ChIP assays using anti-H3K27me3, anti-H4K20me3, anti-H3K9Ac, or anti-H3K4me3 antibody, followed by DNA extraction and real-time PCR to detect DNA fragments using the indicated primers.