Extrinsic Apoptosis Pathway Altered by Glycogen Synthase Kinase-3β Inhibitor Influences the Net Drug Effect on NSC-34 Motor Neuron-Like Cell Survival
Effect of the glycogen synthase kinase-3β (GSK-3β) inhibitor VIII on early/late apoptosis in NCS-34 cells under serum withdrawal conditions. (a) NSC-34 cells were incubated in serum-deprived media with or without the GSK-3β inhibitor at the indicated doses for 60 h. Harvested cells were then stained with an Annexin V-FITC kit and were applied to a fluorescence-activated cell sorting analysis. Early apoptotic cells were Annexin V-positive (right lower). (b) Quantitative representation of cells in early apoptosis. No difference in the proportion of early apoptotic cells was detected among the four GSK-3β inhibitor VIII-treated groups. Presented as mean (% of all cells counted) ± standard error (SE). (c) NSC-34 cells in late apoptosis were indirectly assessed by detecting the change in cleaved caspase-3 signaling by Western blot analysis. Actin was used as the loading control. (d) Quantitative immunoreactivity data of cleaved caspase-3, expressed in arbitrary units and normalized to the control. Reduced cleaved caspase-3 signals were noted in the low-dose GSK-3β inhibitor VIII-treated group but the signal increased significantly in the 1000 nM GSK-3β inhibitor treatment. (compared with control under serum deprivation only). (compared with 200 nM GSK-3β inhibitor-treated group).
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