Glucagon agonist induces β-catenin stabilization in GCGR-expressing cells.

<p>A). Lanes 1–3, HEK293 cells cultured in 6-well plate were transfected with 2000 ng GCGR for 24 h and then either left non-treated(NT) or treated with 50 nM of glucagon agonist (GCG1-29) or antagonist (GCG9-29) for 1 h. Lanes 4–5, HEK293 cells cultured in 6-well plate were transfected with 2000 ng pcDNA3.1 empty vector for 24 h and then treated without or with 20 mM LiCl for 1 h. The cells were harvested and lysed, and equal amounts of protein for each sample were used for western blot analysis. The blot was first probed with anti-β-catenin antibody, then stripped and reprobed for anti-β-actin antibody as a loading control. B). Hep3B hepatocarcinoma cells were serum starved for 4 h and then treated with 50 nM GCG1-29 for the indicated time in serum-free medium. The cells were harvested and lysed. Cell lysates were used for western blot analysis. C). Mouse primary hepatocytes were treated without or with 50 nM GCG1-29 for 1 h and cells were harvested and lysed for western blot analysis.