2 7 dichlorofluorescin diacetate dcfh da

HT (purity ≥98%) was obtained from Extrasynthese (Lyon, France). Dulbecco's modified Eagle's medium (DMEM) was from BiochromAG (Berlin, Germany). RNA was isolated using the RNeasyPlus Mini kit (Qiagen, Hilden, Germany) and cDNA was prepared with Maxima First Strand cDNA Synthesis Kit for RT-qPCR (Fermentas Intl., Vilnius, Lithuania). Real-time PCR was performed using SYBR Fast Master Mix (2x) Universal (KAPABiosystems, Massachusetts, USA). Primers were synthesized by Biomedal S.L. (Sevilla, Spain). Nrf2 siRNA (sc-37049), scramble siRNA (scr siRNA) and the transfection reagent were from Santa Cruz Biotechnology (CA, USA). Opti-MEM reduced serum medium was bought from Thermo Fisher Scientific. α-Tubulin antibody, foetal bovine serum (FBS), sulforhodamine B (SRB), trichloroacetic acid (TCA), 2´,7´-dichlorofluorescin diacetate (DCFH-DA), glutathione reductase (GR), reduced glutathione, NADPH, cumene hydroperoxide, and other general reagents were from Sigma (St. Louis, MO, USA). Heme oxygenase-1 (HO-1) was measured with the enzyme linked immunosorbent assay (ELISA) kit CSB-E08266h from Cusabio (Barksdale, USA). Primary antibodies (PGC-1α, Nrf2, ERRα, SIRT3) were purchased from Santa Cruz Biotechnology, Inc., (CA, USA) except anti-α-tubulin (Sigma, St. Louis, Mo, USA).

Indocyanine green (ICG), poly(D,L-lactide-co-glycolide) (PLGA, MW, 5000–15000; lactide, glycolide (50:50)), hematoxylin and eosin, Sodium sulfide (Na₂S), and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (USA). Bovine hemoglobin (Hb) was purchased from J&K Scientific Ltd. (China). Soybean lecithin, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-maleimide (polyethylene glycol 2000) (DSPE-PEG) were obtained from Avanti (USA). Coomassie Blue, Calcein-AM and Propidium Iodide (PI) Cell Apoptosis Kit were obtained from Invitrogen (USA). Fetal bovine serum, dulbecco’s modified eagle medium (DMEM) and penicillin-streptomycin were purchased from Gibco Life Technologies (USA). 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium (MTS) was bought from Promega (USA). Amicon ultra-4 centrifugal filter with a molecular weight cutoff of 100 kDa was bought from Millipore (USA).

RPMI 1640, phosphate buffered saline (PBS), and yeast peptone dextrose broth (YPD) were obtained from (Thermo Fisher Scientific, Waltham, MA, USA); 3-N-morpholinopropanesulfonic acid (MOPS) was obtained from (Merck); propidium iodide (PI), 2′,7′-dichlorofluorescin diacetate (DCFH-DA), potato dextrose broth (PDB), sabouraud dextrose agar (SDA), sabouraud dextrose broth (SDB), amphotericin B (AFB), acridine orange (OA), ethidium bromide (EB), and crystal violet used in this study were obtained from Sigma-Aldrich, United States; glacial acetic acid was obtained from Carlo Erba Reagents, Italy; and Fluconazole (FLZ) was obtained from Pfizer.

ACS-grade solvents (acetone, methanol, and 95% ethanol), HPLC-grade solvents (water, methanol, and acetonitrile), glass wool, Gibco fetal bovine serum, glacial acetic acid and hydrochloric acid were acquired from the Fisher Scientific Co., LLC (Suwanee, GA, USA). Sephadex LH-20, Amberlite XAD-16, Folin & Ciocalteu’s phenol reagent, gallic acid (>99%), (+)-catechin hydrate (>99%), chlorogenic acid (≥98%), protocatechuic acid (≥97%), vanillic acid (≥97%), fluorescein 3′,6′-dihydroxyspiro[isobenzofuran-1[3H]9′[9[H]xanthen]-3-one), Trolox [(±)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, 97%], TPTZ (2,4,6-tripyridyl-S-triazine, ≥99.0%), quercetin-3-O-rutinoside hydrate (≥94%), iron(II) sulfate heptahydrate (≥99.0%), iron(III) chloride hexahydrate (97%), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, ≥97%), and 2,2′-azobis [2-amidinopropane] dihydrochloride (ABAP) were purchased from the Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Cyanidin-3-O-glucoside chloride was obtained from the Indofine Chemical Company, Inc. (Hillsborough, NJ, USA). Advanced DMEM, phosphate-buffered saline (PBS), Hanks’ balanced salt solution (HBSS), penicillin-streptomycin, and trypsin-EDTA were purchased from Life Technologies (Grand Island, NY, USA). Human colorectal adenocarcinoma (Caco-2) cells were procured from the American Type Culture Collection (ATCC, Manassas, VA, USA).