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The cells were then stained with DAPI, mounted on a slide, and imaged using a confocal microscope with 40 (oil) immersion objective (Zeiss LSM 980, Carl Zeiss)

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Protein adducts are formed due to binding of 4-HNE and MDA amino acid residues Cys, His and Lys in neuronal proteins such as synaptic and cytoskeleton proteins, ion channels and receptors and mitochondrial enzymes