2026-07-05T15:05:44Z https://researchdata.tuwien.ac.at/oai2d

oai:researchdata.tuwien.ac.at:mdh8d-77d32 2024-08-02T13:30:37Z openaire_data

4.3 TUW.TUDATA 10.48436/mdh8d-77d32 oai:researchdata.tuwien.ac.at:mdh8d-77d32 Mikula, Hannes Hannes Mikula TU Wien Goldeck, Marion Marion Goldeck 0000-0003-1899-4763 TU Wien TU Wien 2024 bioorthogonal chemistry click chemistry click-to-release trans-cyclooctene 2024-07-31 2024-08-02 eng 10.1002/chem.202203069 10.48436/rae3t-35174 1.0.0 Creative Commons Attribution 4.0 International The provided data includes all fluorescence microscopy images (embedded in a pptx-file) as published in 10.1002/chem.202203069 (freely accessible). The images are arranged according to the performed experiments and fluorescence channels and can be exported from the pptx-file in 2048x2048px resolution to external tools. The chemical structures of all compounds are specified in 10.1002/chem.202203069.   Technical information HT1080 cells were seeded into a 96-well plate at 10,000 cells per well and allowed to grow overnight. The medium was removed, and the cells were treated with a 50 nM solution of sulfo-dcTCO-DMEDA-CA4 (compound 15) in media. In situ click-to-release was initiated by addition of DMT (compound 9) at a final concentration of 10 µM. As controls, cells were left untreated or incubated with either the parent drug CA4 (50 nM) or 10 µM DMT (compound 9). After an incubation time of 6 h cells were stained with SiR-tubulin (a fluorogenic, cell permeable and highly specific probe for microtubules). An 11X stock solution of the probe was directly added to the growth medium to obtain a final concentration of 1 µM and incubation was carried out for 1 h. Subsequently, the medium was removed, and cells were stained with Hoechst 33342 nuclear dye (Invitrogen, 5 µM in growth medium) for 10 minutes and washed once with PBS. Multichannel imaging of the cells was carried out in FluoroBrite DMEM medium (Gibco) on an Olympus IX82 microscope.   Summary of results Cell imaging via fluorescence microscopy (scale bars: 50 µm) upon staining with Hoechst 33342 (blue, nuclei) and SiR-tubulin (red, microtubules) shows comparable depletion of tubulin signals after 6 h treatment with CA4 (50 nM) or bioorthogonal activation of prodrug 15 (50 nM) by in situ reaction with DMT (compound 9). No significant change (compared to untreated cells) was observed after treatment with DMT (compound 9) or prodrug 15. FWF Austrian Science Fund 013tf3c58 I 4623 N Bioorthogonal time-controlled intramitochondrial elimination