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. 2023 Jun 29;3(7):1952–1964. doi: 10.1021/jacsau.3c00198

Figure 6.

Figure 6

Thyclotide exhibits cell uptake enhancement compared with aegPNA. (A) Schematic of the fluorescein-labeled (FI) aegPNA 10a and fluorescein-labeled thyclotide 10b–10f used in cell uptake experiments. B* represents thf residue, and X represents 2-aminoethoxy-2-ethoxy acetic acid (AEEA linker). (B) 3D volume reconstruction of 0.15 μm z-steps super-resolution microscopy of an HCT116 cell treated with 2.5 μM 10a for 3 h. The cell was stained with the Membrite Fix 640/660 membrane marker and Hoechst 33342 for nucleus staining. The membrane (red) and nucleus (blue) channels were successively removed from the middle and right pictures, respectively, to better show the cytoplasmic and nuclear diffusion of the aegPNA 10a (green). (C) 3D volume reconstruction of 0.15 μm z-steps super-resolution microscopy of an HCT116 cell treated with 2.5 μM 10f for 3 h. The cell was stained with the Membrite Fix 640/660 membrane marker and Hoechst 33342 for nucleus staining. The membrane (red) and nucleus (blue) channel were successively removed from the middle and right pictures, respectively, to better show the cytoplasmic and nuclear diffusion of the thyclotide 10f (green). (D) FACS data of HCT116 comparing the fluorescence of cells not treated or treated with 10a for 3 h. (E) FACS data of HCT116 comparing the fluorescence of cells not treated or treated with 10f for 3 h. (F) Mean of fluorescence intensity for negative control, aegPNA-treated cells, and thyclotide-treated cells. (G) FACS statistics of aegPNA-treated cells and thyclotide-treated cells. Cells were considered positive for cell uptake when the fluorescence was superior to the fluorescence observed for negative control (non-treated cells).