Figure 4.

Ankrd2 decreases the Gsk3β protein expression level and inhibits its activity. (a) Heat map of gene expression values for Ankrd2 and Gsk3β in proliferating (stage 1), fusing (stage 2), and differentiated (stage 3) primary myoblasts derived from Ankrd2 KO and WT mice infected with AdAnkrd2 or control AdGFP (n=3). (b) Representative western blot showing Gsk3β protein and phosphorylation levels at the same stages as in (a). (c) Western blot analysis for Ankrd2 and Gsk3β protein and phosphorylation levels on lysate from proliferating and differentiating C2C12 myoblasts infected with AdAnkrd2 or AdGFP. Densitometry analysis for total and phosphorylated Gsk3β normalized to β-actin protein levels was performed. (d) Western blot and relative densitometry analysis of active and total Gsk3β normalized to β-actin protein levels from proliferating and differentiating C2C12 cells transfected or not with WT or Ser99Ala mutant (S99A) Ankrd2. MB, proliferating myoblasts, MT, differentiating myotubes at 1 and 6 days (d) after induction. Data are represented as mean±S.D. (n=3). *P<0.05; **P<0.01