Figure 5
From: Hepatic STAMP2 decreases hepatitis B virus X protein-associated metabolic deregulation
Knockdown of STAMP2 accelerates HBx-mediated metabolic phenotypes. (A) HepG2-HBx cells were co-transfected with siNegative control or siSTAMP2. Total RNA was prepared from cells, and mRNA levels were assessed by RT-PCR (upper) and Western blotting (bottom). (B) HepG2-HBx cells were transiently transfected with siNegative control or siSTAMP2. Oil-Red O staining was performed with spectrophotometric quantification of lipid staining at 500 nm. (C) HepG2-HBx cells were transiently transfected with siNegative control or siSTAMP2. The mRNA levels of lipogenic (ACC1, FAS, SCD1, and SREBP1) and adipogenic (aP2 and PPARγ) genes were assessed by RT-PCR. (D) HepG2-HBx cells were transiently transfected with siNegative control or siSTAMP2, followed by incubation with different concentrations of insulin for 1 h. Whole cell lysates were subjected to immunoblotting for PI3K-p85, phospho-PI3K-p85 (Tyr458), Akt1/2, and phospho-Akt (Ser473). (E) HepG2 cells were transiently transfected with HA or HA/HBx in the presence or absence of siSTAMP2. Western blotting was performed using anti-IRS1, anti-STAMP2, anti-HBx, and anti-actin antibodies.
