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Fig. 2. NRADD expression in mouse tissues. (A) Characterization of an anti-NRADD antibody. 293 cells were transfected with full length p75^NTR and NRADD FLAG-tagged at the N-terminus. Cell lysates from N2a or transfected 293 cells were analysed by three identical western blots probed with anti-NRADD, antigen blocked anti-NRADD and anti-FLAG antibodies. The N2a and 293 lanes were exposed to film for 5 minutes and the transfected 293 lanes were exposed for $~$ 30 seconds. (B) Mouse tissues were homogenized and separated into membrane (M) and soluble (S100) fractions. An aliquot of each fraction was deglycosylated in vitro with PNG-F or Endo H. Deglycosylation converted the fuzzy mNR band into a darker dglNR band. sNR indicates the small NRADD isoform. N.S. indicates a non-specific band in the brain lysates that was not blocked by preincubation of the antibody with antigen.

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