FIGURE SUMMARY
Title

Intracellular delivery and deep tissue penetration of nucleoside triphosphates using photocleavable covalently bound dendritic polycations

Authors
Ma, J., Wehrle, J., Frank, D., Lorenzen, L., Popp, C., Driever, W., Grosse, R., Jessen, H.J.
Source
Full text @ Chem Sci
Fig. 1

Designed molecule to enable cellular uptake and spatiotemporal control of the release of nucleotides.

Fig. 2

(A) HPLC analysis of the uncaging of DEAC450 caged ATP 6 at 490 nm. Concentration: 100 μM. Volume: 1 mL. (B) Uncaging kinetics of DEAC450 caged ATP 6 at 490 nm.

Fig. 3

Microscopy images of HeLa cells incubated with 50 μM of compounds from a super resolution Lattice SIM microscope. Scale bar represents 10 μm.

Fig. 4

(A) Colocalization analysis of compounds (G1–NH2–) 19, (G2–NH2–) 21, (G2–guanidine–) 22 with Golgi stain — BODIPY® TR ceramide. Green dots indicate fluorescence generated from compounds. Red color indicates Golgi. The orange color indicates colocalized pixels. (B) Colocalization analysis of compounds (G2–NH2–) 21, (G2–guanidine–) 22 with Golgi stain — BODIPY® TR ceramide at different time points. Green dots indicate fluorescence generated from compounds. Red color indicates Golgi. The orange color indicates colocalized pixels. (C) Colocalization analysis of compounds (G1–NH2–) 19, (G2–NH2–) 21, (G2–guanidine–) 22 with LysoTracker® Deep Red. Green dots indicate fluorescence generated from compounds. Purple dots indicate lysosomes. The white color indicates colocalized pixels. HeLa cells were incubated with 50 μM of compound in the dark at 37 °C for 5 h. Scale bar for the overview represents 10 μm. Scale bar for the inset represents 1 μm. Colocalization indicated as percentage.

Fig. 5

(A) Images of the heads of zebrafish embryos in tissue penetration experiments. Different cross-sections of one embryo for 21 and 22 are shown. Incubation time: 24 h. (B) Images of the heads of zebrafish embryos in tissue penetration experiments. Different cross-sections of one embryo for 21 and 22 are shown. Incubation time: 48 h. Final concentration of applied compounds: 50 μM. Green signals indicate either weak autofluorescence (see (a) blank) or the fluorescence generated from compounds. Scale bar represents 40 μm. (C) Transgenic zebrafish with Tg(sox2: sox2-E2A-QF2)m1517 and Tg(QUAS: nls-mCardinal)m1637 with red fluorescent nuclei in neural stem and progenitor cells were used and treated with 6, 21 and 22 respectively. Incubation time: 48 h. Final concentration of applied compounds: 50 μM. Temp.: 28.5 °C. Scale bar represents 20 μm.

Scheme 1

(A) Synthesis of clickable version of DEAC450 3. (B) Synthesis of DEAC450 caged ATP 6. (a) ETT, DMF, r.t., 5 min. (b) mCPBA, DMF, 0 °C, 5 min. (c) 5–10% (v/v) piperidine, r.t., 40 min, in DMF. The product is obtained as triethylammonium (TEA) salt after purification. (C) Synthesis of dendronized DEAC450 caged ATPs with different transporters attached and structures of dendrons used in this study.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Chem Sci
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