Ahr sirna s1200

Negative control siRNA or AHR siRNA (s1200) (both purchased from Invitrogen) was diluted with RNase-free water (Thermo Fisher Scientific), further mixed with Opti-MEM^TM I Reduced Serum Medium (Thermo Fisher Scientific) and Lipofectamine^TM RNAiMAX Transfection Reagent (Thermo Fisher Scientific), and incubated for 20 min at room temperature. HaCaT cells were seeded in 12-well plates and mixed with siRNA-Lipofectamine complex (final siRNA concentration: 10 nM). After 48 h of incubation, cells were treated with DMSO (0.1%), BaP (1 μM) or BAI (25 μM) for 6 h and collected for RNA extraction. The knockdown efficiency of AHR at 48 h after transfection was determined by qRT-PCR and Western blotting.

Negative control siRNA, AHR siRNA (s1200), and PXR siRNA (s16911) (all purchased from Invitrogen, Carlsbad, CA) were each transfected into HaCaT cells using HiPerFect Transfection Reagent (Qiagen) or Lipofectamine RNAiMAX (Themo Fisher Scientific), in accordance with the manufacturers’ instructions. Cells were mixed with siRNA, incubated for 48 h, and used for scratch assay or western blotting. The knockdown efficiency was assessed by qRT-PCR and western blotting.