Wild type ORFV002 and its mutants in OFTu cells prevent NF-κB p65 (Lys³¹⁰) acetylation.

<p>A. OFTu cells were co-transfected with plasmid pNF-κB p65 and either pEGFP-N1 or pORFV002-EGFP. At 24 hours post transfection, cells were treated with 20 ng/ml of TNF-α for 0, 15, 30 and 60 minutes and harvested at the indicated times. Total cell lysis protein extracts (50 μg) were loaded in 10 percent SDS-PAGE gel, blotted, and probed with antibodies against NF-κB p65 (Lys³¹⁰) (top), NF-κB p65 (middle), or GAPDH (bottom). B. OFTu cells were co-transfected with plasmid pNF-κB p65 and ORFV002 mutants. At 24 hours post transfection, cells were treated with 20 ng/ml of TNF-α for 60 minutes. Total cell lysates were detected by WB similar to A. C. Densitometry of NF-κB-p65 (acety-Lys³¹⁰) bands normalized to loading control GAPDH (**, <i>P<</i>0.005). The results are representative of three independent experiments.