A preliminary study on the interaction between Asn-Gly-Arg (NGR)-modified multifunctional nanoparticles and vascular epithelial cells

Liu, Chunxi; Liu, Tingxian; Yu, Xiaoyue; Gu, Yizhu

The plasmid pCMV-EGFP (pEGFP-N1), which carries the gene of enhanced green fluorescent protein (EGFP) under a cytomegalovirus (CMV) promoter, was propagated in Escherichia coli and purified by an Endo Free Plasmid Maxi Kit (Qiagen, Hilden, Germany). The purity and concentration of pDNA was then measured by a NanoDrop UV-Vis Spectrophotometers (ND-2000C, Thermo, USA). A phycoerythrin (PE)-conjugated anti-human CD13 monoclonal antibody (clone WM15) was purchased from BD Biosciences (USA). A DyLight 488-labelled anti-human caveolin 1 monoclonal antibody (7C8) (NB100-615G) was purchased from Novus Biologicals (USA). Hoechst33342 was purchased from Invitrogen by Life Technologies (USA). Methyl-β-eyclodextfin (MβCD), cholesterol, TritonX-100 and bovine serum albumin (BSA) was purchased from Sigma-Aldrich (USA). All other reagents were of special commercial grade and used without further purification. Deionized water was used throughout the experiment. All other reagents of biochemical and molecular biology grades were obtained from Sigma-Aldrich (China). Doxorubicin (DOX) was purchased from Dalian Meilun Biology Technology Co., Ltd. (Dalian, China).