Fig 2.

Specific inhibition of IFN-α-induced LMP-1 mRNA upregulation by NF-κB inhibitors piceatannol and the Syk inhibitor BAY613606. (A) Fold induction of LMP-1 and ISG56 mRNA expression normalized to GAPDH, quantified by real-time RT-PCR in Daudi cells preincubated for 45 min without inhibitor (no inh) or with 10 μM PD98059 (PD), 5 μM SB203580 (SB), 1 μM wortmannin (WM), 10 μM SP600125 (SP), 5 μM Gö6850 (Gö) (upper panels), 2 μM BAY117082 (BAY), 5 μM MG132 (middle panels), 50 μM piceatannol (PIC), vehicle control (dimethyl sulfoxide [DMSO]), or 500 nM BAY613606 (SYK) (lower panels) and then left untreated or treated with 20 ng/ml IFN-α for an additional 90 min. Primers are listed in Table 1, and PCR conditions are described in the text. (B) Immunoblot analysis of phospho(Tyr701)–STAT-1, phospho(Tyr690)–STAT-2, phospho(Tyr705)–STAT-3, and phospho(Tyr694)–STAT-5 (all antibodies obtained from Cell Signaling) expression in total cell extracts of Daudi cells preincubated for 45 min without (−) or with (+) vehicle control (DMSO) or 50 μM PIC and then left untreated (−) or treated with 10 ng/ml IFN-α (+) for 20 min.