Figure 6

Detection of Rtm expression in transfected 293T cells and infected GSM cells using a Rtm-specific peptide antiserum. A and B, Specificities of rabbit anti-Rtm peptide antibodies. 293T cells were transfected with Env (pKEnv) plasmid (A), or with either parental (pKCR3), Rev (pKcRev), or Rtm (pKcRtm and pKRtm) plasmids, as indicated (B). Cells were radiolabeled 5 h with [³⁵S]-methionine 48 h after transfection. Lysates were subjected to immunoprecipitation and fractionated on an SDS-10% polyacrylamide gel. Immunoprecipitations were performed using either anti-CD™ antibodies, a serum from CAEV-infected goat (anti-CAEV), or anti-Rtm peptide antibodies (anti-Rtm). C, Immunoprecipitation of the Rtm protein from infected cells. GSM cells were either mock infected (-) or infected with CAEV-Cork strain (+). When cytopathic effects appeared in infected cell culture, cells were radiolabeled 5 h with [³⁵S]-methionine, and lysates were immunoprecipitated with either anti-Rtm or anti-CD™ antibodies, as indicated. Immunoprecipitated proteins were resolved on a SDS-15% polyacrylamide gel.