All reagents including As(III), As(V) and DMAs(V) were purchased from Sigma-Aldrich (Mainland, China) unless otherwise mentioned. MAs(V) was purchased from Sunlida (Nanjing, China). MAs(III) was prepared as previously described (Yoshinaga & Rosen, 2014 (link)). TMAs(V)O was obtained as a gift from Dr. Guo-Xin Sun (Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, China).
As 3
Manufactured by Merck Group
Sourced in United States
As(III) is a laboratory reagent used for analytical and experimental purposes. It is a compound containing the trivalent form of arsenic.
Automatically generated - may contain errors
Lab products found in correlation
Digitonin, by Merck Group (2 mentions)
Glycerol, by Genview (2 mentions)
Glucose 6 phosphate (g6p), by Solarbio (2 mentions)
Eagle s minimal essential medium, by Thermo Fisher Scientific (2 mentions)
β mercaptoethanol, by Dingguo (2 mentions)
Bovine serum albumin (bsa), by Genview (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Flavin adenine dinucleotide, by Merck Group (2 mentions)
L glutamine, by Solarbio (2 mentions)
Fetal bovine serum (fbs), by Gemini Bio (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Agilent Technologies (1 mentions)
Synergy 2 plate reader, by Agilent Technologies (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Kim 1, by Boster Bio (1 mentions)
Na k atpase, by Santa Cruz Biotechnology (1 mentions)
Chromatographic grade methanol, by Merck Group (1 mentions)
β actin, by Bioworld Technology (1 mentions)
Didox, by MedChemExpress (1 mentions)
Naphthylethylenediamine, by Sinopharm (1 mentions)
Sulfanilamide, by Sinopharm (1 mentions)
8 protocols using as 3
1
Arsenic Species Acquisition Protocol
2
MTT Assay for Toxicity Evaluation
3
Generating Haploid Yeast Strains with Genetic Modifications
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Haploid, asexual BY4741 cells (MATa; his3Δ1; leu2Δ0; met15Δ0; ura3Δ0) (Brachmann et al, 1998 ), stored at −80°C in 20% glycerol, were used as WT and to initiate founder populations. Gene duplication events were mimicked by transforming WT cells with centromeric URA3 and KANMX4 plasmids from the MoBY collection (Ho et al, 2009 ). Each plasmid contained a single gene (BY4741 alleles). Point mutations were individually reconstructed in BY4741 backgrounds using in vivo site‐specific mutagenesis, as described (Stuckey et al, 2011 ). Strains were cultivated in synthetically complete (SC) medium containing: 0.14% yeast nitrogen base (YNB, CYN2210, ForMedium), 0.50% ammonium sulphate, 0.077% complete supplement mixture (CSM, DCS0019, ForMedium), 2.0% (w/v) glucose and pH buffered to 5.8 with 1.0% (w/v) succinic acid and 0.6% (w/v) NaOH. Except for glucose, all required nutrients were present in excess. Where indicated, the medium was supplemented with 3 or 5 mM NaAsO2 (As(III), Sigma‐Aldrich) or other environmental challenges as described in Appendix Table S1 .
4
Evaluating Cellular Responses to Oxidative Stress
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Sulforaphane (SF), menadione, digitonin, As(III), and flavin adenine dinucleotide were purchased from Sigma-Aldrich (MO, USA). Cycloheximide (CHX), 4′, 6-diamidino-2-phenylindole (DAPI), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), bromophenol blue, glycerol, and bovine serum albumin (BSA) were obtained from Genview (TX, USA). Glucose-6-phosphate dehydrogenase and nicotinamide adenine dinucleoide phosphate (NADP) were purchased from Regal (Shanghai, China). β-Mercaptoethanol was obtained from Dingguo (Beijing, China). Eagle's minimal essential medium (MEM), and RPMI1640 were acquired from Gibco (CA, USA). Fetal bovine serum (FBS) was purchased from Gemini Bio-product (CA, USA). Glucose-6-phosphate and L -glutamine were obtained from Solarbio (Beijing, China).
5
Induction of tRNA Fragmentation
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For induction of tRNA fragmentation, cultured cells were incubated in media supplemented with 0.5 mM inorganic sodium meta-arsenite (As[III], Sigma) prior to experimental manipulation (2 h for size exclusion, or 1 h for stress granule core enrichment, or 1 h for northern blotting of total RNA).
6
Adsorption of Arsenic on Graphene Oxide
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A standard solution of As(III) (1000 mg/L As(III) in 2% hydrochloric acid, Sigma-Aldrich) was used to prepare a solution with 25 mg/L of As, from which 10 ml were taken and placed in a flask with 0.0125 g of GO. Once the pH was adjusted to 7 the flasks were placed in the black box described previously, this time one of the flask was covered with aluminum foil to avoid UV irradiance on it. The adsorption experiment was carried at 24, 48, 72, 96 and 120 hours under constant magnetic stirring. Finally, all the solutions were centrifuged for 15 minutes at 3500 rpm and then filtered with a 0.45 and 0.2 μm poliethersulfone (PES) membrane (Whatman). Two drops of HNO3 (66.3%, J.T. Beaker) were added to preserve the solution, according to the ASTM D2972-15 Norm for subsequent As quantification.
7
Quantification of Heavy Metals and Transporters
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AGNH pill (weighted 3 g for each), cinnabar (containing 96% of HgS), as well as realgar (containing 90% of As4S4) were all provided by Guangzhou Bai-Yun-Shan Zhong-Yi Pharmaceutical Company Ltd. (Guangzhou, China). Methanol (Chromatographic grade) was purchased from Sigma Chemical Company (St. Louis, MO, USA), and ammonium carbonate (99.999%) was purchased from Aladdin Industrial Corporation (Shanghai, China). The AsIII, AsV, and Hg standard solutions (1000 mg/L) were purchased from Sigma-Aldrich company, using as the standard curve solution. The production of ultrapure water (UPW) at 25°C depended on the Milli-Q® lab water purifying system (Merck, MA, USA). Specific antigens for the primary antibodies were as follows: GSTα1, GSTmu, OCT1, OCT2, MRP1, aquaporin-9 (AQP-9), and β-actin (Bioworld Technology Co, Ltd., MN, USA); Na+-K+-ATPase, and MT-1 (Santa Cruz, CA, USA); KIM-1, GSTpi, MRP2, MRP4, P-gp, and OAT1 (Boster Biological Technology Co, Ltd., Hubei, China).
8
Biochemicals and Reagents for Cell Experiments
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Sulforaphane (SF), menadione, digitonin, As(III), and flavin adenine dinucleotide were purchased from Sigma-Aldrich (MO, USA). Didox was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Cycloheximide (CHX), 4′,6-diamidino-2-phenylindole (DAPI), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), bromophenol blue, glycerol, and bovine serum albumin (BSA) were purchased from Genview (TX, USA). β-Mercaptoethanol was obtained from Dingguo (Beijing, China). Eagle's minimal essential medium (MEM) and RPMI 1640 were acquired from Gibco (CA, USA). Fetal bovine serum (FBS) was purchased from Gemini Bio Products (CA, USA). Glucose-6-phosphate and L -glutamine were obtained from Solarbio (Beijing, China). Glucose-6-phosphate dehydrogenase and nicotinamide adenine dinucleotide phosphate (NADP) were obtained from Regal (Shanghai, China). Solvents used for extraction and isolation were of analytical grade and obtained from Tianjin Fuyu Fine Chemical Company (Tianjin, China). Naphthylethylenediamine and sulfanilamide were obtained from Sinopharm Chemical Reagent Co. Ltd. (China).
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