Fig. 1
From: Vascular-targeted TNFα and IFNγ inhibits orthotopic colorectal tumor growth
Purification, targeted delivery and functional characterization of IFNγ and TCP-1/IFNγ. a Schematic representation of TCP-1/IFNγ and IFNγ fusion proteins. The TCP-1 peptide was fused to N-terminal of IFNγ protein. b Purification of TNFα, TCP-1/TNFα, IFNγ and TCP-1/IFNγ. Recombinant proteins were purified using Ni–NTA resin followed by SDS-PAGE and coomassie blue staining. c Activity analysis of IFNγ and TCP-1/IFNγ on L929 and Colon 26 cells. Cell viability was determined by MTT assay. d 50 nmol IFNγ or TCP-1/IFNγ was i.v. injected into tumor-bearing mice. Mice were sacrificed 1 h later and localization of IFNγ or TCP-1/IFNγ was detected by anti-His tag antibody (green). White arrows indicate areas where TCP-1/IFNγ was colocalized with CD31 (red) in the tumor blood vessels. IFNγ alone did not bind to the blood vessels of tumor tissues. e Immunohistochemistry of MHC-I (H-2Kb). 5 μg IFNγ or TCP-1/IFNγ was i.v. injected into tumor-bearing mice. Mice were sacrificed 24 h later. f 5 μg IFNγ or TCP-1/IFNγ was i.v. injected into tumor-bearing mice. Mice were sacrificed 1 h later. Apoptotic cells (green) in tumor mass were detected by TUNEL assay. TCP-1/IFNγ could obviously induce more apoptosis of tumor cells when compared with the non-targeted IFNγ