The Anti-Inflammatory Activity of HMGB1 A Box Is Enhanced When Fused with C-Terminal Acidic Tail

Figure 2

Western blot and SDS-PAGE analysis of the C tail-fused A box proteins. (a) Identification of the two fusion proteins expressed in E. coli DH5𝛼 by western blot using a mouse anti-human HMGB1 monoclonal antibody directed against A box. Lane 1:A box+C tail; Lane 2:A box+(Gly₄Ser)₃+C tail. (b) Purity analysis of the two fusion proteins by SDS-PAGE. After purification by Ni²⁺-NTA chromatography, the purities of the two fusion proteins were analyzed with SDS-PAGE. Equal amounts of purity proteins (10 𝜇g) were loaded in Figure 2(b), separated by 12% PAGE gel electrophoresis. Lane 1:Molecular weight standards; Lane 2:A box+C tail; Lane 3:A box+(Gly₄Ser)₃+C tail.

(a)

(b)