Fig. 1. Mature NRADD is a 55 kDa glycoprotein. (A) Pulse chase. Full-length NRADD was transiently transfected into 293 cells labeled 24 hours later with ³⁵S-methionine and [³⁵S]cysteine for 15 minutes and chased with cold medium as indicated. Cell lysates were immunoprecipitated with anti-NRADD antibody and analysed by SDS-PAGE and fluorography. `TUN.' indicates sample derived from cells labeled and chased in the presence of tunicamycin (10 µg ml^-1). (B) In vitro deglycosylation. Lysates from 293 cells transfected with NRADD were immunoprecipitated as in (A) and subjected to endoglycosidase digestions as indicated. dglNR, deglycosylated NRADD; iNR, immature NRADD; mNR, mature NRADD.