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2 protocols using sodium dichromate

1

Mechanistic Insights into Chromium-Induced Autophagy

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Rapamycin, sodium dichromate (Na2Cr2O7·H2O) [Cr(VI)], 2,3-diaminonaphthalene (DAN), mercury(II) chloride (HgCl2), sodium hydroxide (NaOH), NO inhibitor aminoguanidine (AG), p38 inhibitor SB203580, NO donor diethylamine NONOate sodium salt, Bafilomycin A1, Chloroquine diphosphate salt, and S-nitrosocysteine were obtained from Sigma–Aldrich (St. Louis, MO). The fluorogenic caspase-9 substrate, LEHD–amino-4-methylcoumarin (AMC), was from Alexis Biochemicals (San Diego, CA). Diaminofluorescein (DAF)-diacetate (DA) was purchased from Molecular Probes (Eugene, OR). Antibodies for Rabbit IgG, Bcl-2, p62, Beclin-1, Atg5, Atg12, P-p38/p38, β-actin, and peroxidase-labeled secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). Lipofectamine 2000 was purchased from Life Technologies (Carlsbad, CA). The Bcl-2 inhibitor ABT-737 and a Bcl-2 antibody for immunoprecipitation were obtained from Santa Cruz Biotechnologies (Dallas, TX). pAb anti-LC3 antibody (HRP) was from Novus Biologicals (Littleton, CO).
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2

Spin Traps for Radical Detection

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Four spin traps were selected to cover a large range of lipophilicity and reactivity against the main families of radicals. DEPMPO and EMPO were obtained from Radical Vision (Marseille, France). POBN and PBN were purchased from Enzo-Alexis Biochemicals (Lausanne, Switzerland). Spin traps were dissolved in water, except for PBN which was dissolved in DMSO. Stock solutions (2 M) were stored at -80°C.
Chemicals for the radical generating systems (xanthine, xanthine oxidase, diethylenetriaminepentaacetic acid (DTPA), ferrous ammonium sulfate (Fe (NH4)2(SO4)2), hydrogen peroxide (H2O2), sodium sulfite, sodium dichromate, sodium azide and horseradish peroxidase) were purchased from Sigma Aldrich (St Louis, MO).
Toxic agents were chosen in order to produce radicals representative of the main types commonly observed (O, N, C or S-centered radicals). Menadione bisulfite, hydrogen peroxide, tert-butylhydroperoxide, phenylhydrazine, catalase (CAT) and superoxide dismutase-polyethylene glycol (PEG-SOD) were obtained from Sigma Aldrich (St Louis, MO).
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