Mammary tissues (100 mg) were homogenized in TRIzol reagent. The total RNA was extracted using the RNeasy lipid tissue kit
(Qiagen, Valencia, CA) and RNA (5 μg) was reverse transcribed with Invitrogen’s SuperScript® III First-Strand
Synthesis System. RT-qPCR was performed using TaqMan rat CYP1A1, CYP1B1, and ACTB primers with FAM/MGB probe (Applied Biosystems,
Carlsbad, CA) as described previously (29 (link)). MCF-10A cells were obtained from ATCC
(Manassas, VA) and authenticated via STR profiling (Promega). MCF-10A cells were cultured as described previously (29 (link)). For the in vitro CYP1A1/CYP1B1 induction experiments, cells having approximately
15 through 20 passages were plated in 96-well plates and treated with vehicle (DMSO), GG, GI, and licorice compounds for 24 h.
RT-qPCR was performed as previously described (29 (link)) using TaqMan® 1-Step RT-PCR
Master Mix, and CYP1A1 and CYP1B1 primer with FAM-MGB probe and GAPDH primer with VIC-MGB probe. Data were analyzed with the
comparative CT (ΔΔCT) method and expressed as fold induction
relative to the vehicle control group.
(Qiagen, Valencia, CA) and RNA (5 μg) was reverse transcribed with Invitrogen’s SuperScript® III First-Strand
Synthesis System. RT-qPCR was performed using TaqMan rat CYP1A1, CYP1B1, and ACTB primers with FAM/MGB probe (Applied Biosystems,
Carlsbad, CA) as described previously (29 (link)). MCF-10A cells were obtained from ATCC
(Manassas, VA) and authenticated via STR profiling (Promega). MCF-10A cells were cultured as described previously (29 (link)). For the in vitro CYP1A1/CYP1B1 induction experiments, cells having approximately
15 through 20 passages were plated in 96-well plates and treated with vehicle (DMSO), GG, GI, and licorice compounds for 24 h.
RT-qPCR was performed as previously described (29 (link)) using TaqMan® 1-Step RT-PCR
Master Mix, and CYP1A1 and CYP1B1 primer with FAM-MGB probe and GAPDH primer with VIC-MGB probe. Data were analyzed with the
comparative CT (ΔΔCT) method and expressed as fold induction
relative to the vehicle control group.
