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Adenoviruses were produced by cotransfection of the PacI-digested pacAd5 U6-shRNA and pacAd5 9.2-100 plasmid into Adeno-X 293 cells (Clontech Laboratories, CA)

Oxidative damage in keratinocytes exposed to cigarette smoke and aldehydes

.,GutteridgeJMHalliwellBThe antioxidant proteins of extracellular fluids

KEAP1 mass spectrometry analysis For mass spectrometry analysis, 10 g of recombinant human KEAP1 (#11981-H29B-100, Sino Biologicals, Beijin, PR China) were diluted to 100 l with 20 mM Tris pH 8.0 and diafiltrated by means of a Amicon Ultra-0.5 ml device (Sigma-Aldrich, Milan, Italy) to eliminate GSH, and then treated with 30 mM M DTT and incubated 30 min at room temperature to reduce the disulfide bridges, diafiltrated to remove the excess of DTT and diluted in 20 mM Tris pH 8.0

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Glutathione peroxidases, as well as other enzymes, rapidly convert two GSH to GSSG, while reducing and thereby detoxifying various hydroperoxides