FIG 4.

RNA bridges the interaction between VDAC1 and IBDV VP1-VP3 complexes. (A) 293T cells were transfected with VP1 and/or VDAC1 expression plasmids. Cell lysates were prepared 48 h after transfection, and then lysates with VDAC1-VP1 association were subjected to RNA digestion with RNase A and RNase T₁ for 1 h at room temperature. Untreated cells were used as controls. All lysates were used for immunoprecipitation (IP). IP was performed with anti-Myc antibody, and lysates were immunoblotted with anti-FLAG or anti-Myc antibody. (B) 293T cells were transfected with VP3 and/or VDAC1 expression plasmids, and lysates of the VDAC1-VP3 cotransfection mixture were subjected to RNA digestion with RNase A and RNase T₁ for 1 h at room temperature. Untreated cells were used as controls. All lysates were used for immunoprecipitation (IP). IP was performed with anti-HA antibody, and lysates were immunoblotted with anti-FLAG or anti-HA antibody. (C) A portion of each sample was analyzed for RNA content by electrophoresis on a 1% agarose gel. Lanes: −, samples without RNA digestion; +, samples with RNA digestion.