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2 protocols using sodium dichromate na2cr2o7.2h2o cr 6

1

Pharmacological Modulation of Cellular Pathways

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Amino guanidine (AG), Thiazolyl blue tetrazolium bromide (MTT), Cisplatin, Rapamycin, Bafilomycin A1, Sodium dichromate (Na2Cr2O7.2H2O) (Cr[VI]), Pan-caspase inhibitor zVAD.FMK, and Akt-inhibitor LY294002 were obtained from Sigma-Aldrich (St Louis, MO). Mn(III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) and Bcl-2 inhibitor VI (ABT-737) were purchased from Calbiochem (La Jolla, CA). The oxidative probes, 4,5-Diaminofluorescein diacetate (DAF-2DA), Dichlorofluorescein (DCF) diacetate, Dihydroethidium (DHE), and the apoptosis dye Hoechst 33342 were from Molecular Probes (Eugene, OR). Antibodies for Beclin-1, p38 MAPK, GAPDH, p-Akt Ser 473, total-Akt, NF-κB, and peroxidase-labeled secondary rabbit and mouse antibodies were from Cell Signaling Technology. Antibody for superoxide dismutase (SOD) was procured from Sigma Aldrich. Antibodies for Bcl-2 and LC3 were from Santa Cruz Biotechnology and Novus Biologicals, respectively. Cyto-ID® autophagy detection kit was purchased from Enzo Life Sciences. Bicinchoninic acid (BCA) was obtained from Thermo Scientific and chemiluminescent substrate from Pierce (Supersignal® West Pico, Pierce).
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2

Evaluation of Cellular Stress Responses

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Thiazolyl blue tetrazolium bromide (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)), cisplatin, sodium dichromate (Na 2 Cr 2 O 7 .2H 2 O) (Cr(VI)), Triton X-100, DNAse-free RNAse, and propidium iodide were obtained from Sigma Aldrich (St. Louis, MO). The oxidative probes 4,5-diaminofluorescein diacetate (DAF-2DA), dichlorofluorescein (DCF) diacetate, dihydroethidium (DHE), and the apoptosis dye Hoechst 33342 were obtained from Molecular Probes (Eugene, OR). Antibodies for PARP, caspase-9, p-Akt (Ser 473), total-Akt, p27, cyclin D3, cyclin-dependent kinase 2 (CDK2), p21, nuclear factor kappa B (NF-κB), and IkappaB kinase-beta (IKK-β) and peroxidase-labeled secondary rabbit and mouse antibodies were purchased from Cell Signaling Technology (Denvers, MA). Antibody for Bcl-2 was procured from Santa Cruz Biotechnology (Dallas, TX). Bicinchoninic acid (BCA) and SuperSignal ® West Pico chemiluminescent substrate were purchased from Thermo Fisher Scientific (Waltham, MA).
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