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In brief, 200 mg of fresh seedlings were flash frozen and ground to a fine powder using sterile wooden pestles and vortexed in three volumes of extraction buffer containing 100 mM Tris-HCl (pH 7.5), 100 mM NaCl, 5 mM EDTA, 5% SDS, 20% glycerol, 20 mM DTT, 40 mM β-mercaptoethanol, 2 mM phenylmethylsulfonyl fluoride, 40 µM MG115 (Apexbio Technology LLC, Houston, TX), 40 µM MG132 (Cayman Chemical, Ann Arbor, MI), 1× phosphatase inhibitor cocktail 2 (Thermo Fisher Scientific), 1× phosphatase inhibitor cocktail 3 (MilliporeSigma), 1× EDTA-free protease inhibitor cocktail (MilliporeSigma), and 0.01% bromophenol blue

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Continue at your current tirzepatide dose and simply use the B6 formulation for your next vial