Figure 4
AhR triggers G β /PI3K/Akt/NF-κB signaling. (A) The PI3K and Gβ protein interaction was assessed by immunoprecipitation. Huh7 cells were immunoprecipitated with antibodies against Gβ or PI3K. Normal IgG was used as a negative control. (B) Cells were treated with BBP (1 μM) for various durations, and then PI3K and phosphorylation levels of Akt were determined by immunoblotting. (C) Nuclear and cytoplasmic fractions of NF-κB were detected by immunoblotting. Lamin A/C and α-tubulin were used as internal markers for nuclear and cytoplasmic proteins, respectively. Huh7 cells were pretreated with wortmannin (100 nM) and then treated with BBP (1 μM) for 2 hours. (D) The Akt phosphorylation levels were measured and β-actin was used as an internal control. (E) The nuclear and cytoplasmic fractions of NF-κB were analyzed by immunoblotting. Huh7 cells were transfected with two different shRNA and treated with BBP (1 μM) for 2 hours. (F) PI3K, p-Akt, Akt levels were measured by immunoblotting. β-actin was used as an internal control. (G) The nuclear and cytoplasmic fractions of NF-κB were analyzed by immunoblotting. Histone H3 and α-tubulin were used as internal markers for nuclear and cytoplasmic proteins, respectively.