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4 protocols using dipyridamole dipy

1

Glucuronidation Profiling of Flavonoids

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Alamethicin, β-estradiol, dipyridamole (DIPY), D-saccharic-1, 4-lactone, magnesium chloride (MgCl2), MK-571, Ko143, leukotriene C4 (LTC4) and uridine diphosphate glucuronic acid (UDPGA) were all obtained from Sigma-Aldrich (St Louis, MO). Apigenin, chrysin, cycloicaritin, genistein, and wushanicaritin with purity over 98% was purchased from Aladdin Reagents (Shanghai, China). Apigenin-7-O-glucuronide, chrysin-7-O-glucuronide, genistein-7-O-glucuronide, and β-estradiol-3-O-glucuronide was purchased from Toronto Research Chemicals (North York, ON, Canada). Wushanicaritin-3-O-glucuronide was prepared in our laboratory as described previously [23 (link)]. 293T cells, HeLa cells, pLVX-mCMV-ZsGreen-PGK-Puro vector [9371 base pairs (bp)] and pLVX-shRNA2-Neo vector (9070 bp) were all provided from BioWit Technologies (Shenzhen, China). The pGEM-T plasmid which carried the UGT1A1 cDNA clone was purchased from Sino Biological Inc. (Beijing, China), while recombinant human UGT1A1 was purchased from Corning Biosciences (New York, USA). The anti-BCRP, anti-GAPDH, anti-MRP1, anti-MRP2, anti-MRP3, anti-MRP4 and anti-UGT1A1 antibodies were all obtained from OriGene Technologies (Rockville, MD). All other chemicals and reagents were the highest grade commercially available.
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2

Comprehensive Analytical Reagent Protocol

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Ammonium acetate, sodium chloride (NaCl), 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 2-mercaptoethanol, benzamidine, protease inhibitor cocktail, N-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK), phenylmethanesulfonyl fluoride (PMSF), adenosine 5′-triphosphate (ATP), amino propyl trimethoxy silane (APTS), gluteraldehyde aqueous solution, avidin, N-(+)-Biotinyl-6- aminohexanoic acid (Biotin-X), dipyridamole (DIPY), mesoporphyrin IX (MIX), protoporphyrin IX (PIX), PK-11195 (PK), rotenone (Rot), flunitrazepam (Flu), glipizide, glibenclamide and diclofenac were obtained from Sigma-Aldrich (St. Louis, MO, USA or Munich, Germany), tris(hydroxymethyl)aminomethane (Tris) and glycerol were obtained from Applichem (Darmstadt, Germany), ethylenediaminetetraacetic acid (EDTA) was obtained from Scharlau (Barcelona, Spain). Dialysis tubing was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Open tubular capillaries (100 μm i.d.) were obtained from Polymicro Technologies (Phoenix, AZ, USA). De-ionized water was obtained from a Milli-Q system (Millipore, Molsheim, France). All other chemicals used were of analytical grade.
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3

Lentiviral Vector Construction and Characterization

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The pLVX-mCMV-ZsGreen-PGK-Puro vector [9371 base pairs (bp)], and a pLVX-shRNA2-Neo vector (9070 bp) were obtained from BioWit Technologies (Shenzhen, China). Expressed human UGT1A1 was purchased from Corning Biosciences (New York). Alamethicin, β-estradiol, β-estradiol-3-O-glucuronide, dipyridamole (DIPY), D-saccharic-1,4-lactone, magnesium chloride (MgCl2), MK-571, Ko143, leukotriene C4 (LTC4), and uridine diphosphate glucuronic acid (UDPGA) were purchased from Sigma-Aldrich (St Louis, MO). UGT1A1, GADPH, BCRP, MRP1, MRP3, and MRP4 antibodies were all purchased from OriGene Technologies (Rockville, MD). Bisdemethoxycurcumin was provided from Guangzhou Fans Biotechnology Co., Ltd (Guangzhou, China). All other chemicals and reagents were of guaranteed reagent or the highest grade commercially available.
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4

Corylifol A Metabolism Assay

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Corylifol A (purity >98%) was purchased from Shanghai Winherb Medical Technology Co., Ltd (Shanghai, China). Pooled human liver microsomes (HLM), nine individual human liver microsomes (iHLM), mice liver microsomes (MLM), monkeys liver microsomes (MkLM), rabbits liver microsomes (RaLM), rats liver microsomes (RLM), dogs liver microsomes (DLM), twelve recombinant human UGT isoforms (UGT1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 2B4, 2B7, 2B10, 2B15 and 2B17) and twelve expressed human CYP enzymes (CYP1A1, 1A2, 1B1, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4 and 3A5) were all obtained from Corning Biosciences (Corning, NY, USA). The anti-BCRP (TA322704), anti-MRP1 (TA309559), anti-MRP3 (TA314800) and anti-MRP4 (TA327332) antibodies were purchased from OriGene Technologies (Rockville, MD). UDPGA, NADPH, MgCl2, alamethicin and D-saccharic-1, 4-lactone, dipyridamole (DIPY) and MK571 were all provided from Sigma-Aldrich (St Louis, MO, USA). Paclitaxel, 6-hydroxy-paclitaxel, β-estradiol and propofol were supplied from Aladdin Chemicals (Shanghai, China). Propofol-O-glucuronide and β-estradiol-3-O-glucuronide were obtained from Toronto Research Chemicals (Ontario, Canada). Besides, other chemicals and solvents were of analytical grade or better.
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