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. 2012 Jun 18;33(7):888–896. doi: 10.1038/aps.2012.73

Figure 4.

Figure 4

Vam3 inhibits the activation of FoxO3a, which is increased by CSC in Beas-2B cells. Beas-2B cells were treated with CSC (300 mg/L) for 12 h, with or without RES (1 μmol/L, 0.1 μmol/L) or Vam3 (1 μmol/L, 0.1 μmol/L) pretreatment for 2 h. (A) Cell extracts were immunoprecipitated with an anti-FoxO3a antibody, and the immune complexes were subjected to immunoblotting analyses with anti-acetyl-lysine antibody. IgG was used as the negative control. (B) The amounts of acetylated FoxO3a were calculated by densitometry and normalized to corresponding FoxO3a levels. (C) Equal amounts of cytoplasmic (cytoplasm) or nuclear (nucleus) protein extracts (60 μg/lane) were loaded and the membranes were probed with an anti-FoxO3a antibody. GAPDH served as the cytoplasmic control and LaminB was used as the nuclear control. (D) The relative intensity of nucleus/cytoplasm was calculated by densitometry. Mean±SD. n=3. cP<0.01 vs control; eP<0.05, fP<0.01 vs that in the absence of RES or Vam3.