Fig. 8.

Inhibition of SIRT2 by AGK2 as well as silencing of SIRT2 in combination with Lapatinib upregulates acetylated FOXO3 levels in 5-8F and 6-10B cells. a NPC were treated with AGK2 alone or in combination with Lapatinib for 48 h. Proteins were obtained from whole cell extracts and assessed by co-immunoprecipitation (co-IP) with an anti-FOXO3 antibody. Subsequent immunoblotting was performed using antibodies against Ac-Lys and FOXO3. The anti-IgG was used as a negative control. Acetylated FOXO3 levels were determined by using ImageJ analysis and representative bar diagram are shown (bottom panel after the blot). b NPC cells were transiently transfected with siNSC and siSIRT2 and treated with 5 μM Lapatinib for 24 h. Proteins were obtained from whole cell extracts and assessed by Co-immunoprecipitation (Co-IP) with an anti-FOXO3 antibody. Subsequent immunoblotting was performed using antibodies against Ac-Lys and FOXO3. Acetylated and total FOXO3 levels were determined by using ImageJ analysis and representative bar diagrams demonstrating relative acetylated to total FOXO3 ratios are shown (bottom panel after the blot).