(a)
E. coli BL21 [DE3] cya cells carrying pET23::AC, where AC was either IlaC* (IlaC22 k27 Y259F), IlaD9 or IlaM5, and a plasmid expressing heme oxygenase, pT7-ho112. Cells were collected post-induction with IPTG, as described in Material and Methods. Loaded are whole cell extracts (W), soluble (S) and Insoluble (I) fractions. Red arrows point to the position of the NIRW-AC protein bands. (b) HEK293 cells transfected with equal amounts of either pAAV::ilaC* or pAAV::ilaM5 DNA and grown for 24 h prior to collection. Equal amounts of total soluble protein from each culture were analyzed using SDS-PAGE. Left panels, NIRW-AC protein abundance assayed via Western blotting using a His6-specific antibody. Right panel, β-actin-specific antibody used to verify equal protein loading. Molecular mass markers (kD) are shown on the sides of each panel. n.t., not transfected.