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. 2015 Oct 15;8(10):17565–17577.

Figure 1.

Figure 1

Plasmid DNA vaccines encoding secreted scFvN418-HER2/neu fusion proteins. A. Schematic representation of expression vectors. scFvN418-HER2, scFvN418-neu, pcDNA3.1-HER2, or pcDNA3.1-neu encode under the control of a CMV promoter, all the fusion proteins consisting of an signal peptide, amino acid residues 1 to 222 of human HER2 or amino acid residues 1 to 224 of rat neu, and COOH-terminal Myc tag. The control plasmids pcDNA3.1-HER2 and pcDNA3.1-neu lack the scFvN418 domain. B. 293T cells grown in 100-mm dishes were transfected with various expression vectors using Lipofectamine 2000 (invitrogen). Immunoblot analysis of supernatants from 293T cells transfected with scFvN418-HER2, scFvN418-neu, pcDNA3.1-HER2 and pcDNA3.1-neu (lane 1, 2, 3 and 4). Vaccine proteins were probed with mouse anti-Myc tag mAb followed by HRP-conjugated secondary anti-mouse antibody. C. Binding of scFvN418-HER2 fusion protein from culture supernatant of transfected 293T cells to mouse dendrtic cells was investigated by fluorescence-activated cell sorting analysis with mAb 9E10 followed by PE-conjugated secondary antibody. Control cells were treated with supernatant from mock-transfected cells.