Hela human cervix adenocarcinoma cells

HT29 human colorectal adenocarcinoma cells, HeLa human cervix adenocarcinoma cells, and HDFa human adult dermal fibroblasts as control cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The IGROV1 human ovarian cancer cell line was kindly provided by Prof. Colnaghi (Istituto Nazionale Tumori, IRCCS, Milan, Italy). Cells were cultured in RPMI 1640 medium (Labtek Eurobio, Milan, Italy), and supplemented with 10% FCS (Euroclone, Milan, Italy) and 2 mM L-glutamine (Sigma-Aldrich, Milan, Italy), at 37 °C, and a 5% CO₂ atmosphere. The complexes were dissolved in DMSO in a 30–40 mM stock solution. In cell treatments, the final DMSO concentration never exceeded 0.1%.

H4 human neuroglioma cells, HeLa human cervix adenocarcinoma cells, and MDA-MB-231 human mammary gland adenocarcinoma cells were obtained from the American Type Culture Collection (Manassas, VA). H4 and HeLa cells were maintained in Dulbecco's modified Eagle's medium (DMEM), and MDA-MB-231 in DMEM-F12 (ThermoFisher, Waltham, MA). For all cell lines, media were supplemented with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin (ThermoFisher), and 5 μg/ml plasmocin (InvivoGen, San Diego, CA), and cells were cultured in a humidified incubator with 5% CO₂ at 37 °C. For transient transfections, cells were either seeded on top of glass coverslips on 24-well plates or on 6-well plates. When cells were ∼60% confluent, transfections were performed with Lipofectamine 2000 (ThermoFisher), according to the manufacturer's instructions. Preparation of protein extracts from cultured, transfected, or nontransfected cells was performed using methods described elsewhere (Tenorio et al. 2016 ).

HeLa human cervix adenocarcinoma cells, SH-SY5Y human neuroblastoma, and HDFa human normal fibroblast as control cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in RPMI 1640 medium (Labtek Eurobio, Milan, Italy), supplemented with 10% FCS (Euroclone, Milan, Italy) and 2 mM L-glutamine (Sigma-Aldrich, Milan, Italy), at 37 °C, and a 5% CO₂ atmosphere. γ-T, α-T, p-cym, and myr were purchased from Sigma-Aldrich (Sigma-Aldrich, Milan, Italy). γ-T, α-T, p-cym and myr were percolated twice through activated basic alumina and once through silica to remove impurities and traces of hydroperoxides. The compounds were dissolved in DMSO in a 30 mM stock solution. In cell treatments, the final DMSO concentration never exceeded 0.1%.