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Potassium Ferrate(VI) Preparation and Characterization

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Carbamazepine (CBZ, >98% purity),
sulfadimethoxine (SDM, >98% purity), trimethoprim (TMP, >98%
purity),
all test amino acids (i.e., glycine, alanine, serine, leucine, proline,
phenylalanine, glutamic acid, asparagine, and histidine), phenyl methyl
sulfoxide (PMSO), phenyl methyl sulfone (PMSO2), sodium
thiosulfate, and sodium borate were purchased from Fisher Scientific
(Austin, Texas, USA). A wet chemical procedure was applied to prepare
solid potassium ferrate(VI) (K2FeO4),59 (link) which had a purity of more than 90%. The solid
K2FeO4 was dissolved in a 0.01 M sodium borate
buffer solution. The concentration of Fe(VI) solution was determined
by measuring the absorbance at 510 nm using a 1.0 cm path length and
a molar absorption coefficient of ε510nm = 1150 M–1 cm–1,59 (link) with a UV–visible spectrometer (Thermo Scientific Co., USA).
All reaction solutions were prepared using Milli-Q ultrapure water
(>18 MΩ cm–1 resistivity, Millipore, Milford,
USA).
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2

Lysosome-related Organelle Regulation

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Hank’s balanced saline solution (HBSS), pyruvate, and L-glutamine were purchased from Corning. Chloroquine (CQ) was purchased from MP Biomedicals. Bafilomycin A (baf A), leupeptin, and Torin 1 were purchased from Cayman chemical. Carbonyl cyanide m-chlorophenylhydrazone (CCCP) was purchased from Enzo Life Sciences. Bortezomib was purchased from LC Laboratories. Pepstatin, glycine, lysine and serine were purchased from Sigma. Glucose, asparagine, and histidine were purchased from Fisher. Arginine was purchased from Acros Organics. The siRNAs against TFEB, TFE3, and SNAT7(siGenome SMARTpool) and the control scramble siRNA were purchased from Dharmacon. Rabbit antibodies used in this study were against LC3B (Abcam), P62, S6K, pT389-S6K, ATG5, ATG7, TFEB, TFE3 (Cell Signaling Technology), SNAT7 (Sigma), and Limp2 (Novus Biologicals). Mouse antibodies used in this study were against GAPDH (Developmental Studies Hybridoma Bank (DSHB)), Lamp1 [38 (link)](DSHB), and CD63 [39 (link)] (DSHB). TFEB-EGFP was a gift from Shawn Ferguson (Addgene plasmid #38119) [10 (link)]. HRP-conjugated secondary antibodies for Western blotting were purchased from Jackson Laboratory. Secondary antibodies conjugated to Alexa Fluor were purchased from Molecular Probes.
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