Purification of recombinant mNT5E. (A) Diagram of the GST-mNT5E expression construct. (Top) Native mNT5E contains an N-terminal signal peptide (ss-cleavage) and GPI anchor site. (Bottom) The GST-mNT5E fusion construct contains the signal peptide from gp67 of baculovirus Autographica californica, GST, a thrombin cleavage site, the catalytic domain of mNT5E and (His)6 tag. Translation start and stop codons are indicated. (B) GelCode blue-stained SDS-PAGE gel and (C) western blot of purified recombinant mNT5E protein (0.05 μg). The western blot was probed with an anti-mNT5E antibody.