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2 protocols using 4 hydroxy trans 2 nonenal 4 hne

1

Oxidative Stress and Cellular Pathways

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4-Hydroxy-trans-2-nonenal (4-HNE) was purchased from Cayman Chemical (Ann Arbor, MI). Luteolin (≥98%), apigenin (≥97%), chrysin (≥97%), MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide), RPMI-1640 medium, H2O2, N-acetyl-cysteine (NAC), tunicamycin (Tm), protein kinase A (PKA) inhibitor H-89 [N-[2-((p-Bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide] and other chemicals were purchased from Sigma-Aldrich Co. (St. Louis, MO), unless otherwise indicated. SB 203580 (4-[4'-fluorophenyl]- 2-[4'-methylsulfinylphenyl]-5-[4'-pyridyl] imidazole), a p38 MAP kinase inhibitor was purchased from Promega (Madison, WI). MEK inhibitor U0126, and c-Jun NH2-terminal kinase (JNK) inhibitor SP600125 (anthra[1,9-cd]pyrazole-6 (2H)-one) were purchased from Calbiochem (San Diego, CA).
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2

Lipid Peroxidation and Membrane Biophysics

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1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1-oleoyl-2-hydroxy-sn-glycero-3-phosphocholine (OPC), bovine heart cardiolipin (CL), arachidonic acid (AA), hexane, hexadecane, KCl, HEPES, EDTA, Na2SO4, MES, Tris, adenosine 5-triphosphate (ATP), and guanosine 5-triphosphate (GTP) were purchased from Sigma-Aldrich (Munich, Germany). MPC (1-myristoyl-2-hydroxy-sn-glycero-3-phosphocholine) was obtained from Avanti Polar Lipids (Alabaster, AL, USA). Plain silica beads (22-μm diameter) were from Microspheres-Nanospheres (Corpuscular Co., Cold Spring, NY, USA). Chloroform was purchased from Carl Roth (Karlsruhe, Germany). 4-oxo-trans-2-nonenal (4-ONE) and 4-hydroxy-trans-2-nonenal (4-HNE) were purchased from Cayman Chemical (Ann Arbor, MI, USA) or synthesized as previously described [45 (link)].
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