Hexachloronaphthalene Induces Mitochondrial-Dependent Neurotoxicity via a Mechanism of Enhanced Production of Reactive Oxygen Species

Figure 4

PCN67 treatment induces characteristics of necrotic death in differentiated PC12 cells. (a) Representative dot plots of necrotic/apoptotic cell distribution following 72 h of PCN67 treatment. Cells were stained with Annexin V and propidium iodide and analyzed with flow cytometry. (b) Quantification of viable (V), early apoptotic (EA), late apoptotic (LA), and necrotic (N) cells in a population. (c) Determination of plasma membrane integrity measured by the release of lactate dehydrogenase (LDH) 72 h following PCN67 treatment. (d) Western blot-based quantification of HMGB1 protein in cytosolic fraction collected from 72 h-treated cells. The results are presented as arbitrary units following normalization to the GAPDH level used as a marker of a cytosolic fraction. Histone H3, a protein marker of a nuclear fraction, was used to determine the purity of fractionation. (e) Average measurement of the nuclear diameter of differentiated PC12 cells after 72 h of PCN67 treatment. (f) The changes in the ATP level following 72 h of PCN67 treatment. The results were normalized to the protein level and are expressed as nmoles/mg. and .

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