Figure 1
NCS-induced glioma cell death is caspase-9 dependent and involves mitochondria. (a) Viability of glioma cells treated with different concentration of NCS for 24 h as determined by MTS assay. (b) Viability of glioma cells treated with 1 μg/μl NCS in the presence and absence of TNFα (50 ng/ml) for 24 h as determined by MTS assay. (c) Western blot of cleaved caspase-9, Bad, Bax and Cytochrome c in glioma cells treated with different combinations of NCS and TNFα. A representative blot is shown from three independent experiments with identical results. Blots were reprobed for β-actin to establish equivalent loading. (d) Viability of glioma treated with different combinations of TNFα and NCS in the presence and absence of Caspase-9 inhibitor, as determined by MTS assay. The graph (a, b, d) represents the viable glioma cells percentage of control. * denotes significant change from control (P<0.05). #denotes significant change from NCS and NCS+TNFα (P<0.05). (e) Compromised mitochondrial integrity in NCS-treated cells both in the presence and absence of TNFα, as demonstrated by MitoTracker green staining. Cells treated with different combinations of NCS and TNFα were stained with MitoTracker green FM and examined at × 40 magnification. (f) Intracellular ATP content of glioma cells treated with different combinations of NCS and TNFα.* denotes significant change from control (P<0.05). (g) NCS decreases lactate production in glioma cells both in the presence and absence of TNFα. Graph indicates lactate levels in cells treated with different combinations of NCS and TNFα. Values represent the means±S.E.M. from three independent experiments. #denotes significant change from control (P<0.05). *denotes significant change from TNFα (P<0.05)
