Figure 2.

Ankrd2 represses the transcriptional activity of NF-κB during oxidative stress. (a) C2C12 myoblasts were transfected with the 3 × NF-κB-luc vector and pGL4.74 control vector before infection with AdAnkrd2 or AdGFP. The values represent the firefly/Renilla luciferase ratio in proliferating and differentiating cells. (b) Luciferase activity assay in C2C12 cells following cotransfection of 3 × NF-κB-luc vector with FLAG-tagged WT or Ser99Ala mutant (S99A) Ankrd2. (c) Effect of 4 h of treatment with hydrogen peroxide on NF-κB transcriptional activity in C2C12 myoblasts at 1 day after induction of differentiation. Where indicated, cells were pretreated with Akt1/2- and Akt-specific inhibitors, as previously described.⁸ (d) Western blot analyses on the same samples as in (c), showing increased phosphorylation levels of Akt and Ankrd2. MB, myoblast; MT, differentiating myotubes at 1, 3, and 6 days (d) after induction. Data are represented as mean±S.D. (n=3). *P<0.05; **P<0.01