Figure 6
NCS-induced ATM phosphorylation regulates NFκB and TIGAR. (a) Western blot indicating increased ATM phosphorylation in cells treated with NCS both in the presence and absence of TNFα. (b) ATM regulates NFκB in NCS-treated cells. NFκB transcriptional activity in cells transfected with NFκB luciferase reporter construct and treated with different combinations of NCS, TNFα and ATM inhibitor Ku60019. The graph represents fold change in NFκB luciferase activity over control. Values mean±S.E.M. from three independent experiments. *denotes significant change from control (P<0.05), #denotes significant change from TNFα and NCS+TNFα (P<0.05). Inset shows pATM levels in cells treated with NCS in the presence and absence of ATM inhibitor, as determined by western blot. (c) ATM phosphorylation in NCS-treated cells is independent of NFκB. Western blot demonstrating ATM phosphorylation in cells transfected with IκBM and treated with NCS in the presence and absence TNFα. (d) ATM regulates TIGAR in NCS and TNFα cotreated cells. Western blot demonstrating TIGAR levels in cells treated with NCS or TNFα or both in the presence of KU60019. Figures (a, c, d) are representative of three independent experiments. Blots were reprobed for β-actin to establish equivalent loading. (e) NCS reduces mitochondrial genome content. The graph represents mtDNA copy number per cell by comparing nuclear genes to the corresponding mitochondrial genes, as determined by qRT-PCR. Values represent the mean±S.E.M. from two independent experiments. *denotes significant change from control (P<0.05)
