Coimmunoprecipitation of NF45 and IBDV proteins after RNase treatment. Lysates of pcDNA3-ckNF45-FLAG-transfected and subsequent IBDV-infected DF-1 cells (A) or DF-1 cells only IBDV infected (B) were subjected to RNA digestion with either RNase T 1 or RNase III or double-treated (T1+ R III) for 1 h at 37°C. Untreated cells were used as controls. One portion of each sample was subsequently analyzed for RNA content on a 1% agarose gel (left panel). The remaining portions of the samples were used for immunoprecipitations (IP). IP were performed with either the anti-FLAG MAb (A) or the VP2-specific MAb R63 and the VP3-specific MAb IBDV2 (B). The resulting precipitates were separated by SDS-PAGE and analyzed by Western blotting (WB) using either rabbit anti-ckNF45-His serum (anti-VP3 or -VP1) or IBDV protein-specific antiserum (rabbit anti-IBDV serum).