Fig. 2: PEG-coated DNA nanotube seeds attach to EGFR receptors via AMDA.

a, d Three-dimensional projection images of HeLa (a) and HEK293 (d) cells after AMDA or AMDA with BDC tag addition omitted (No BDC tag) to attach seeds to EGFR (Supplementary Notes S20 and S23). Nanotube seeds were labeled with Atto488 (red) and secondary antibody–streptavidin conjugates with Alexa647 (blue). Scale bars: 20 μm. b Confocal micrograph cross-sections of HeLa cells stained with DiD dye (blue) (Supplementary Note S21) before seeds (red) were attached via AMDA. Scale bar: 20 μm. This experiment was repeated independently more than three times with similar results. c Average fluorescence intensities of nanotube seeds per HeLa cell after AMDA or after AMDA omitting different reagents with 32 pM seeded added (Supplementary Note S22, N = 9 random locations for each case were analyzed). For no 1AB: only the addition of EGFR primary antibody was omitted during the AMDA, no 2AB-STA: only the addition of secondary antibody and streptavidin conjugate (2AB-STA) was omitted, no BDC’ tag on seeds: there were not BDC’ tags modified on nanotube seeds. e Confocal micrograph cross-sections of HEK293 cells. Scale bar: 20 μm. This experiment was repeated independently more than three times with similar results. f HEK293 cell fluorescence in the channel used to label seeds after AMDA (orange), AMDA with BDC tag addition omitted (blue), and no AMDA (red), measured via flow cytometry (Supplementary Note S24). 48 pM seeds were added to cells. Average fluorescence intensities were 1423 ± 75 (N = 9818 cells), 883 ± 55 (N = 9836 cells), and 759 ± 6 (N = 9867 cells). g–i Co-localization of nanotube seeds and antibodies on the cell membrane (Supplementary Note S25). g Confocal micrograph cross-section of a HeLa cell after AMDA with secondary antibody labeled with Alexa647 (green) and nanotube seeds labeled with atto488 (32 pM) (red). Scale bar: 10 μm. h Computerized localization of antibodies (green) and seeds (red) of cells in (g). i Mean fractions of nanotube seeds colocalized with antibody after AMDA and in randomized controls (N = 12 cells were analyzed). Error bars here in (c), (f), and (i) are 95% confidence intervals.