Nicotine

Human ovarian cancer cell lines: SKOV3 (HTB-77), TOV112D (CRL-11731), and TOV21G (CRL-11730) were purchased from ATCC (American Type Culture Collection). The HEY ovarian cancer cell line was a kind gift from Dr. Jinsong Liu. SKOV3, TOV112D, TOV21G, and HEY cells were cultured at 37 °C with 5% CO₂ in DMEM media supplemented with 10% calf serum (CS), penicillin and streptomycin. Cells were treated with a final concentration of 0.1 mM, 0.5 mM, or 2 mM nicotine (MP Biomedicals, Inc) for 24 h-96 h. After the treatments, the cells were counted and whole cell extracts were prepared.

Nicotine was purchased from MP Biomedicals, Inc. (Solon, OH). Diethyl ether was purchased from Nacalai Tesque (Kyoto, Japan). Trizol reagent and primers were obtained from Invitrogen (Carlsbad, CA). ReverTra Ace qPCR RT master Mix and THUNDERBIRD SYBR qPCR Mix were obtained from Toyobo (Tokyo, Japan). Complete mini protease inhibitor was purchased from Roche Applied Science (Vilvoorde, Belgium). Chenodeoxycholic acid (CDCA) was purchased from Wako (Osaka, Japan). UDP-glucuronic acid (UDPGA), alamethicin, and saccharic acid 1,4-lactane were purchased from Sigma—Aldrich (St Louis, MO). Human liver microsomes were obtained from BD Gentest (Woburn, MA). All the other reagents were of the highest grade commercially available.