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AHR knockdown

JG J. Gilbert
GI G. N. De Iuliis
AM A. McCluskey
JS J. A. Sakoff
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Transient knockdown of AHR in MDA-MB-468 cells was performed through transfection of small interfering RNAs (siRNA) targeting AHR (Qiagen) and the AllStars Negative Control nonsilencing siRNA (Qiagen). The AHR siRNA contained four siRNAs for the AHR target (FlexiTube GeneSolution GS196). Cells were transfected with Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions. Briefly, 8 × 103 MDA-MB-468 cells were plated into each well of a 96-well plate and allowed to adhere for 24 h. Opti-MEM media (Invitrogen) containing 0.3 mL of Lipofectamine 3,000 transfection reagent and 0.3 pmol siRNA was added to each well. After 6 h of incubation, transfection media was replaced with growth media containing 0.5 µM NAP-6. Cells were incubated for a further 48 h prior to MTT analysis23.

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