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2 protocols using 5 fluorodeoxyuridine

1

Reagents for Biomolecular Modification

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5-Fluorodeoxyuridine was purchased from Alfa Aesar (Shanghai, China). 4,4ʹ-Dimethoxytrityl Chloride (DMT-Cl, 97%) N,N-Diisopropylethylamine (DIPEA, 97%) and 2-Cyanoethyl N,N,N’,N’-tetraisopropylphosphordiamidite (95%) were obtained from Macklin Inc. (Shanghai, China). N-(ε-malemidocaproyloxy) succinimide ester (EMCS) was purchased from Aladdin (Shanghai, China). Ni-NTA agarose, and Sephadex G-25 were obtained from GE Healthcare (Piscataway, USA). Imidazole, sodium chloride, sodium acetate, polyacrylamide, tris base, acetic acid, ethylenediaminetetraacetic acid (EDTA), magnesium chloride, ethanol, and DAPI were obtained from Sangon Biotech (Shanghai) Co., Ltd (China). Amicon ultracentrifugal filters were purchased from Merck Millipore Ltd (Darmstadt, Germany). RPMI 1640 medium, DMEM medium, MEBM medium, trypsin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), antibiotic-antimycotic (100×) and fetal bovine serum (FBS) were purchased from Wisent Biotechnology (Nanjing) Co. Ltd (China). All chemicals were used without further purification.
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2

Synthesis and Characterization of Fluorescent Probes

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5-fluorodeoxyuridine (98%, Alfa Aesar, Shanghai, China), 4,4’-Dimethoxytrityl Chloride (DMT-Cl, 97%, Macklin, Shanghai, China), N, N-Diisopropylethylamine (DIPEA, 97%, Macklin), 2-Cyanoethyl N, N, N′, N′-tetraisopropylphosphordiamidite (95%, Macklin), N-(ε-malemidocaproyloxy) succinimide ester (EMCS, 98%, Aladdin, Shanghai, China), pyrene (95%, Aladdin), curcumin (95%, Aladdin) and 2-(4-amidinophenyl)-6-indolecarbamindine dihydrochloride (DAPI, 95%, Sangon Biotech, Shanghai, China) were used as received. CytoPainter LysoRed Indicator Reagent (ab176828) was bought from Abcam Co., Ltd (Boston, MA, USA). Trypsin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), phosphate-buffered saline (PBS), Dubelcco’s Modified Eagle’s Medium (DMEM), Roswell Park Memorial Institute-1640 (RPMI-1640), antibiotic-antimycotic (100×) and fetal bovine serum (FBS) were purchased from Wisent Biotechnology (Nanjing) Co. Ltd (Nanjing, China). The caspase 3, 8 and 9 activity detection kits were purchased from BestBio Biotechnology Co., Ltd. (Shanghai, China).
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