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4 protocols using hydroquinone

1

Optimized DNA Amplification Protocol

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Primer STAR HS DNA polymerase (Takara, Dalian, China) was used for PCR. Restriction endonucleases and a DNA purification kit were purchased from Thermo Scientific (Waltham, USA). Plasmid and genomic DNA were extracted using the Plasmid Miniprep Purification kit (Sangon Biotech, Shanghai, China) and the TIANGEN pre kit (Tiangen, Beijing, China), respectively. Oligonucleotide synthesis and sequence analyses were performed by Sangon Biotech (Shanghai, China). Hemin and 5‐aminolevulinic acid hydrochloride (ALA) were obtained from Sigma‐Aldrich (St. Louis, MO, USA). Phenol, hydroquinone, mevastatin, pravastatin sodium, daidzein, and 7,3′,4′‐trihydroxyisoflavone were obtained from Macklin (Shanghai, China). Other chemicals obtained were of the highest commercial grade available from Sangon Biotech (Shanghai, China).
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2

Genomic DNA and plasmid extraction protocol

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PrimeSTAR HS DNA polymerase used for PCR was purchased from Takara (Dalian, China). The kits for genomic DNA and plasmid extraction, and DNA gel purification were purchased from Takara, Sangon (Shanghai, China), and Thermo Scientific (Shanghai, China), respectively. Sangon performed oligonucleotide synthesis and Sanger sequencing. ALA, methanol, and acetonitrile were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Phenol and hydroquinone were obtained from Macklin (Shanghai, China). Other chemicals were purchased from Sangon unless otherwise specified.
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3

Mycotoxin Standardized Immunoassay Protocol

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Tenuazonic acid and other common mycotoxin standards, bovine serum albumin (BSA) and goat anti-mouse IgG antibody (GAMA) were obtained from Sigma-Aldrich (MO, United States). Trisodium citrate, HAuCl4 4H2O and hydroquinone were purchased from Macklin (Shanghai, China). Nitrocellulose (NC) membrane was provided by Jieyi BioTech. (Shanghai, China). Anti-TA antibody reagent (anti-TA McAb 6D5) was prepared in our laboratory according to our previous work (Cai et al., 2021 (link)). All the other reagents were analytical reagent grade. Ultraviolet-visible spectrum (UV-VIS) was tested by spectrophotometer (PerkinElmer, Waltham, MA, United States). Tecnai G2 F30 transmission electron microscope (TEM) (FEI, Hillsboro, OR, United States) was used to scan the diameter and shape of the prepared nanoparticles. The UHPLC-MS/MS was performed by Waters ACQUITY UPLC-XEVO TQ-S MS with ACQUITY UPLC BEH C18 (Waters, Milford, MA, United States).
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4

Zebrafish Compound Screening Protocol

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Calycosin was purchased from Chengdu Gelipu Bio Co., Ltd. (Chengdu, China). Hydroquinone (≥99%, determined by HPLC), arbutin (≥98%), and kojic acid (≥99%) were purchased from a commercial supplier Macklin (Shanghai, China). Ethyl 3-aminobenzoate methanesulfonate was purchased from Aladdin (Shanghai, China). Other regular reagents for zebrafish system maintains were purchased from Wuhan TianZhengYuan Biological Technology Co., Ltd. (Wuhan, China). All buffers and other reagents were of the highest purity commercially available.
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