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. 2019 Feb 27;47(10):e55. doi: 10.1093/nar/gkz137

Figure 2.

Figure 2.

Flow cytometric determination of delivery specificity; Binding and uptake of antibody-Cy3 and DNA-Cy5 (before and after chromatin assembly) was analysed by flow cytometry after incubation for 1 h. (A) Histogram of MCF7 cells after treatment with targeted (anti LeY; dotted red) and untargeted (anti CD33; dotted blue) DNA-Cy5 complexes. Cy5 signal was detected only after treatment with the targeted DNA-Cy5 construct. (B) Histogram of MCF7 cells after treatment with targeted (red) and untargeted (blue) chromatin-Cy5 complexes. Results are comparable to results after DNA–Cy5 delivery. (C) Contours plot of MCF7 Cy3 (x-axis) and Cy5 (y-axis) signals after treatment with various antibody chromatin complexes comprising Cy3 labelled antibody and Cy5 labelled DNA. Cells were treated with complexes comprising antibody without specificity against cell surface antigen but against CPXM2 peptide do neither show Cy3 nor Cy5 signals (blue). Cells treated with complexes comprising digoxigenin CPXM2 peptide (instead of biotin CPXM2 peptide) display Cy3 signal but no Cy5 signal, demonstrating that antibody but not chromatin is present at the cell surface (green). Cells treated with complexes comprising antibody with specificity against the cell surface and CPXM2 peptide display Cy3 signal and Cy5 signal, demonstrating that antibody as well as chromatin is present at the cell surface (red).