4 4 azobis 4 cyanovaleric acid acva

The solvents petroleum ether (40–60 °C), diethyl ether, dichloromethane, ethyl acetate and acetone, and the reagents cyanomethyl dodecyl trithiocarbonate, 4,4′-azobis(4-cyanovaleric acid) (ACVA) and 4-dimethylaminopyridine (DMAP) were purchased from Sigma Aldrich and used as received. N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC·HCl) was purchased from Alpha Aesar and used as received. 1,4-Dioxane, and the monomers DEGMA, OEGMA (M_n = 300 g mol^–1), nBA and DMA were purchased from Sigma Aldrich and passed through a column of basic alumina prior to use. NIPAM was purchased from Sigma Aldrich and recrystallized from a toluene–hexane mixture prior to use. 2,2′-Azobisisobutyronitrile (AIBN) was purchased from Sigma Aldrich and recrystallized from methanol prior to use. Dialysis membrane (MWCO = 3.5–5 kDa) was purchased from Spectra/Por. The synthesis of N,N-diethylacrylamide,⁶¹
2-cyano-2-propyl dodecyl trithiocarbonate,⁶²
4-cyano-4-(((ethylthio)carbonothioyl)thio) pentanoic acid⁶³
have been described previously in the literature.

Dimethyl acrylamide (DMAm, 99 %; Sigma Aldrich 274135), Acrylic Acid (AA, 99 %; Sigma Aldrich 147230), (Methacryloyloxy)ethyl] trimethylammonium chloride (QDMAEMA, 75 % in H₂O; Sigma Aldrich 408107), (Methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl)ammonium hydroxide (DMAPS, 95 %; Sigma Aldrich 537284), 4,4′-azobis(4-cyanovaleric acid) (ACVA, 99%; Sigma Aldrich 11590), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC, 98%; Sigma Aldrich 341006), deuterated methanol (CD₃OD, 99.8%; Sigma Aldrich 151947-10G-GL) and deuterium oxide (D₂O, 99.9%; Sigma Aldrich 151882) were purchased from Sigma Aldrich (UK). DAAm (99%) was purchased from Alfa Aesar (UK; A15940.30). 4,4-Difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acyl hydrazide (BODIPY FL, 99 %) was purchased from ThermoFisher Scientific (UK; D2371). 4-((((2-Carboxyethyl)thio)carbonothioyl)thio)−4-cyanopentanoic acid (BM1433, 95%) was purchased from Boron Molecular (USA; BM1433).
For protoplast preparation, cellulase “Onozuka RS” was purchased from Melford biolaboratories (Duchefa; C8003.0005) and pectolyase from Sigma Aldrich (P3026).

All the following chemicals were used without
further purification: sulfobetaine methacrylate (SBMA, 98%, Taiwan
Hopax Chemicals), 4,4′-Azobis(4-cyanovaleric acid) (ACVA, 98%,
Sigma-Aldrich), 4-cyanopentanoic acid dithiobenzoate (CPAD, 97%, Strem
Chemicals), dopamine hydrochloride (DA, 98%, Tokyo Chemical Industry),
ethanol (95%, ECHO), acetone (99%, ECHO), isopropyl alcohol (IPA,
95%, Uni-Onward), methanol (95%, Uni-Onward), deuterated water (D₂O, 99.9%, Sigma-Aldrich), 2,2,2-trifluoroethanol (TFE, 99.8%,
Acros Organics), oxalic acid (H₂C₂O₄, 97%, Showa), chromium(VI) oxide (CrO₃, 98%, Showa),
copper chloride (CuCl₂, 98% Showa), hydrochloric acid (HCl,
>37%, Honeywell Fluka), phosphoric acid (H₃PO₄, >85% J. T. Baker), and perchloric acid (HClO₄, 70%,
Showa). Deionized (DI) water was obtained by a pure water system (RODA)
with a resistivity of 18.25 MΩ·cm. A 0.1 M tris(hydroxymethyl)aminomethane
(Tris) buffer solution with pH = 8.1 was prepared by dissolving a
preset pHast Pack powder (PPB023, Sigma-Aldrich) in DI water to a
total volume of 500 mL. 0.2 mm-thick aluminum foil (Al foil, 99.997%),
1 mm-thick aluminum oxide (Al₂O₃), and 1 mm-thick
glass substrates were procured from Alfa Aesar, Cheng Yang Instrument
Corp., and FEA, respectively.

ε-Caprolactone (CL, Acros, 99% purity), 2-ethylhexanoic acid tin(II) salt (Sn(Oct)₂; Sigma-Aldrich, St. Louis, MO, USA, purity ≥95%), 4,4′-Azobis(4-cyanovaleric acid) (ACVA; Sigma-Aldrich, purity ≥ 98%), 4-Cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPA; Sigma-Aldrich, purity ≥ 97%), poly(ethylene glycol) methyl ether methacrylate (PEGMA; Sigma-Aldrich, M_n 950 Da), Tween80 (Sigma-Aldrich), Diethyl ether (DEE, Sigma-Aldrich, ≥99%), Deuterated chloroform (CDCl₃, Sigma-Aldrich, 99.8%), Ethanol (EtOH, Sigma-Aldrich, 99.8%), acetonitrile (ACN, Sigma-Aldrich, 99.9%), 6-hydroxycaproic acid (HCA, ABCR, Karlsruhe, Germany, 95%) were used as received without further treatments. The cell medium is composed by high glucose DMEM/F12 (Biowest, Nuaillé, France) supplemented with 10% fetal bovine serum (Lonza, Basel, Switzerland). Phosphate-buffered saline solution (PBS) was purchased from Sigma-Aldrich. For gel permeation chromatography (SEC) analysis Tetrahydrofuran (THF, Sigma-Aldrich, purity ≥ 99.7%) was used as eluent.

Hematoporphyrin dihydrochloride (HP) was purchased from MedChem Express (San Diego, CA, USA). RPMI-1640 medium with L-glutamine and phenol red, methanol, and N-hydroxysuccinimide were purchased from FUJIFILM Wako Pure Chemical (Osaka, Japan). RPMI 1640 medium without phenol red was purchased from Thermo Fisher Scientific (San Jose, CA, USA). RIPA buffer was purchased from Nacalai Tesque (Kyoto, Japan). 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (WSCD-HCl) was purchased from Peptide Institute, Inc. (Osaka, Japan). Trypsin-ethylenediaminetetraacetic acid (EDTA), [2-(methacryloyloxy)ethyl]trimethylammonium chloride (METAC), 4,4′-azobis(4-cyanovaleric acid) (ACVA), and N-(3-aminopropyl)methacrylamide hydrochloride (APMAA) were purchased from Sigma-Aldrich (St Louis, MI, USA). 2,2,6,6-Tetramethyl-4-piperidinol (4-OH-TEMP) was purchased from the Tokyo Chemical Industry (Tokyo, Japan). Fetal bovine serum (FBS) was purchased from Gibco (Waltham, MA, USA). The LIVE/DEAD Cell Staining Kit was purchased from Dojindo (Kumamoto, Japan). The Apoptosis/Necrosis Assay Kit (ab176749) was purchased from Abcam (Cambridge, UK). 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).