Figure 2
NCS-induced NFκB, Akt and Erk, activation regulates prosurvival response in the presence of TNFα. (a) NCS increases TNFα-induced NFκB activation in glioma cells. The graph represents fold change in NFκB luciferase activity over control, in cells treated with TNFα or NCS or both for 24 h. Values represent the means±S.E.M. from three independent experiments. * denotes significant change from control, #denotes significant change from TNFα-treated cells (P<0.05). (b) NCS-mediated cell death in the presence and absence of TNFα is increased in cells transfected with IκBM. Viability of mock transfected or IκBM transfected glioma cells treated with different combinations of TNFα and NCS, was determined by MTS assay. (c) Western blot analysis indicating Akt and Erk phosphorylation in glioma cells treated with TNFα or NCS or both for 24 h. Representative blot is shown from three independent experiments with identical results. Blots were reprobed with β-actin to establish equivalent loading. (d) Treatment with Akt inhibitor enhances NCS-induced glioma cell death. Viability of glioma treated with different combinations of TNFα and NCS in the presence and absence of Akt inhibitor LY294002, as determined by MTS assay. (Inset) Akt inhibitor abrogates pAkt levels in cells treated with different combinations of TNFα and NCS as determined by western blot analysis. The graph (b, d) represents viable glioma cells expressed as percentage of control. Values (b, d) represent the means±S.E.M. from three independent experiments. * denotes significant change from control, #denotes significant change from mock transfected (b) or NCS+TNFα (d) (P<0.05)
