AMWAP is secreted from reactive BV-2 microglia. (A) BV-2 microglial cells were treated with 50 ng/ml LPS for 48 h before obtaining cellular protein lysates (L) as well as concentrated culture supernatants (S). Anti-AMWAP immunoblot analysis revealed a band of approximately 12 kDa only present in the supernatant of LPS-activated microglia. GAPDH and Ponceau S served as loading controls. (B) Anti-AMWAP immunoblot analysis of recombinantly expressed and purified AMWAP from E. coli (AMWAP-His) and HEK293 EBNA cells (AMWAP-Strep(II)) shows single bands at approximately 11 to 13 kDa.