Fig. 6.

Cellular uptake of dual-labeled HyApNc consisting of A550-4 and A647N-3 motifs. a–c Confocal fluorescence images of H1838 cells treated with the HyApNc consisting of Atto550-DxR-L4 motif (A550-4) and Atto647N-trCLN3-L4 (A647N-3) motifs in 1:1 ratio. Both A647N-3 (a: c2: red) and A550-4 (b: c3: magenta) fluorescence were observed from the cytosol including a FRET-mediated Atto647N signal (c: c4: cyan). d Calculated FRET signal from reconstructed FRET images (d: white) indicate the intracellular integrity of the functional nanoconstruct HyApNc. e, f Overlay images of cells incubated with HyApNc (e: A647N-3 + A550-4), and HyApNc.mut (f: A647N.mut-3 + A550-4) as a negative control with Atto647N-labeled mutant trCLN3.mut-L4 motif. The complete overlay sets for e and f are shown in Supplementary Fig. 19. Aptamer constructs were incubated at 37 °C for 2 h, followed by membrane staining with Alexa488-WGA (green), and nuclei staining with Hoechst 33342 (blue). Scale bar: a–f 50 µm