Cell culture reagents

LNCaP human AR+ (ATCC^® CRL-1740™), DU145 (ATCC^® HTB-81™) and PC3 human AR− (ATCC^® CRL-1435™) prostate cancer cell lines were obtained from the American Type Culture Collection (ATCC) (Rockville, MD, USA). They were maintained as previously described [7 (link),8 (link)], and they were tested periodically for mycoplasma contamination using PlasmoTest™—Mycoplasma Detection Kit (InvivoGen, San Diego, CA, USA). Anti-α-tubulin and bovine serum albumin (BSA) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Rat tail collagen and transwell inserts were obtained from BD Biosciences (San Jose, CA, USA). DAPI (4′,6-Diamidino-2-Phenylindole) was purchased from Invitrogen through Thermo Fisher Scientific (Eugene, OR, USA). Rabbit polyclonal anti-Gα_i2 antibody (ab 157204) was purchased from Abcam (Cambridge, MA, USA). Epidermal growth factor (EGF) was obtained from Life Technologies (Grand Island, NY, USA). The anti-rabbit and anti-mouse immunoglobulins coupled with horseradish peroxidase (IgG-HRP), were obtained from Promega (Madison, WI, USA). Cell culture reagents were purchased from Corning Life Sciences (Tewksbury, VA, USA).

Pooled human liver microsomes from 150 donors (75 males; 75 females) were purchased from Corning Life Sciences (Woburn, MA, USA), and (‒)-sophoranone (99.7% purity; SPN) was supplied by SK Chemicals Ltd. (Sungnam, Gyeonggi-do, Korea). β-Nicotinamide adenine dinucleotide phosphate disodium salt (NADP), glucose 6-phosphate disodium salt hydrate, glucose 6-phosphate dehydrogenase, MgCl₂, and all chemicals including the specific substrates, its metabolites, and well-known inhibitors of nine P450s were purchased from Sigma–Aldrich Corporation (St. Louis, MO, USA), Santa Cruz Biotechnology (Dallas, TX, USA), or Cayman Chemicals (Ann Arbor, MI, USA) unless stated otherwise. The purity of all purchased compounds was higher than 97.0%. HPLC-grade acetonitrile and methanol were obtained from Burdick & Jackson Company (Morristown, NJ, USA). Caco-2 cells were supplied by the Korean Cell Line Bank (Seoul, Korea) and cultured according to the supplier’s recommendations. Transwell (24-well, 6.5 mm polycarbonate inserts, 0.4-μm pore) and cell culture reagents were purchased from Corning Life Sciences. Heparinized human plasma was obtained from donors at the Severance Hospital of Yonsei University Health System (Seoul, Korea) and stored at −80 °C prior to use.

Cinnamon powder was purchased from a local distributor. C. cassia was from Carmel Organics, Barukheda, India; C. loureiroi was from Kirkland, D.C, USA; C. zeylanicum was from Sorich Organics, Noida, India. Infliximab was purchased from a local Pharmacy in India. Cell culture reagents were purchased from Corning (NY, USA). All other reagents were purchased from the vendors as listed. Caspase3/7 glo reagent (G8090, Promega Madison, WI, USA), TNF-α (AB155699, Abcam, Waltham, MA, USA), trans-cinnamaldehyde (TCA; C80687, Sigma, Burlington, MA, USA), cinnamic acid (CA; 8.00235, Sigma), eugenol (E51791, Sigma), actinomycin D (A9415, Sigma), trypsin–EDTA (25,200–056, Gibco, Billings, MT, USA), phosphate-buffered saline (PBS), pH 7.2 (20,012–027), and Alamar Blue (DAL1100, Invitrogen, Waltham, MA, USA).

Chemicals (substrates, drugs, assay medium components) were from Sigma-Aldrich (St. Louis, MO) unless otherwise noted. Cell culture reagents were from Corning (Bedford, MA). Seahorse XF consumables (cartridges, cell culture plates) were from Agilent (Santa Clara, CA). The bicinchoninic (BCA) assay for protein, tetramethylrhodamine methyl ester (TMRM) and other fluorescence probes were from Life Technologies (Carlsbad, CA). The FLIPR Membrane Potential Assay Explorer Kit was from Molecular Devices (Sunnyvale, CA; #R8126 red version). Zosuquidar was from Cayman Chemical (Ann Arbor, MI).

SW480 human colon cancer, MCF-7 human breast cancer, SKOV3 human ovarian cancer, HTB-35 human squamous cervical cancer cells and untransformed human HSF2617 and HFF fibroblasts were obtained from the ATCC (Manassas, VA, USA). All cell lines were cultured in DMEM at 37 °C and 5% CO₂ pressure. Medium was supplemented with 10% fetal bovine serum, 4 mM glutamine, 1 mM pyruvate, 10 and 25 mM glucose, and 1% (v/v) penicillin and streptomycin in bicarbonate buffer at pH 7.2. Cell culture reagents were obtained from Corning through ThermoFisher (Fairlawn, NJ, USA).