Gαq 11

Whole cell extracts were prepared in RIPA lysis buffer (Millipore, Temecula, CA, USA) containing 1× protease inhibitor cocktail (Thermo Scientific, Waltham, MA, USA). Protein concentrations were determined using a BCA protein assay kit (Thermo Scientific, Rockford, IL, USA). Equal amounts of protein (50 μg protein) were resolved by sodium dodecyl sulfate-polyacaryamide gel electrophoresis (SDS-PAGE), transferred onto a polyvinylidene difluoride membrane, and blocked with 5% nonfat dry milk for 1 hour at room temperature. After blocking, the membrane was incubated overnight with primary antibodies at 4°C and washed three times with PBST. The horseradish peroxidase conjugated secondary antibodies (Santa Cruz Biotechnology) were incubated for 1 hour at room temperature. The blots were washed three times with PBST and then visualized with an enhanced chemiluminescence kit (Thermo Scientific, Rockford, IL, USA). The primary antibodies were as follows: AhR, Gα_q/11, G_β, PIP2, IP3R, and PI3K from Santa Cruz Biotechnology (1:100); p44/42 MAPK (Erk1/2), Akt, p-Akt (ser473), NFκB, LaminA/C, Histion H3 and α-tubulin from Cell Signaling Technology (1:1000); COX-2 from Abcam (1:1000); β-actin from Sigma (1:1000).