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Fig. 7 . Effect of NP60 on the activation of ATF2. (A) 293T cells were grown on 35 mm dishes. Luciferase reporter plasmid (5xGAL, 2 µg), pRL-SV40 (100 ng), GAL-ATF2 (200 ng), NP60 (200 ng) or MKK6b (200 ng) and FLAG-p38{alpha} (200 ng) were co-transfected into 293T cells in different combinations as indicated. Luciferase activity was measured 24 hours after transfection. Values are the means ± s.e.m. (B) Flag-p38{alpha} was co-expressed with myc-NP60 in 293T cells. Flag-p38{alpha} was isolated by anti-Flag antibody. It was then incubated with ATF2 fusion protein in the presence of ATP. The reaction complex was resolved by SDS-PAGE and western blotting was performed using anti-phospho-ATF2 antibody.





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