Mccoy s 5a media

Human mammary carcinoma cell lines BT-474, SK-BR-3, and MCF-7 cells were purchased from American Type Culture Collection (ATCC). BT-474 cells were maintained in RPMI-1640 media (Cat# SH30027.01, Hyclone, GE Healthcare Life Sciences, Chicago, IL, USA) supplemented with 10% fetal bovine serum (FBS, Cat# SH30071.03HI, Hyclone, GE Healthcare Life Sciences). SK-BR-3 cells were maintained in McCoy’s 5A media (Cat# 16600-082, Life Technologies, Carlsbad, CA, USA) supplemented with 10% FBS. MCF-7 were grown and maintained in minimum essential medium (MEM) (ThermoFisher Scientific, Cat# 42360032, Waltham, MA, USA) supplemented with 10% FBS, non-essential amino acids (NEAA) (ThermoFisher Scientific, Cat# 11140050) at 1X, and recombinant human insulin (ThermoFisher Scientific, Cat# 12585014, Waltham, MA, USA) at 10 µg/mL.

The human teratocarcinoma cell line Tera-1 (ATCC, Manassas, VA, USA, Cat# HTB-105) was grown in McCoy’s 5A media (Life Technologies, Carlsbad, CA, USA, Cat# 16600-082), supplemented with 15% FBS (Atlanta Biologicals, Norcross, GA, USA, Cat# S11195) and 1% Pen-Strep (Life Technologies Cat# 15140-122). Feline astrocyte G355.5 cells (ATCC Cat# CRL-2033) were grown in McCoy’s 5A media supplemented with 10% FBS and 1% Pen-Strep. 293T cells (ATCC Cat# CRL-2316) were grown in DMEM supplemented with 10% FBS and 1% Pen-Strep. All cell lines were grown at 37 °C with 5% CO₂.

HCT116 colon cancer cells were cultured in McCoy’s 5A media (Life Technologies, cat#16600-082, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Life Technologies, cat#10099-141). Genomic DNA was isolated using a Gentra Puregene Cell Kit (Qiagen, cat#158745, Redwood City, CA, USA) as per manufacturer’s instructions. Purified genomic DNA was quantified with a Nanodrop ND-1000 (Thermo Scientific, Carlsbad, CA, USA).

All materials and reagents were purchased from commercial sources and used as received. CPT, dimethyl sulfoxide (DMSO), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and trypan blue were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-buffered saline (PBS), fetal bovine serum (FBS), penicillin/streptomycin, trypLE express enzyme, McCoy's 5A media and Ham's F-12K (Kaighn's) nutrient mix were purchased from Life Technologies (Carlsbad, CA, USA). Paclitaxel was sourced from 21 CEC PX Pharm Ltd., (East Sussex, UK); chloroform and hydrochloric acid, from APS Chemicals (Canning Vale, WA, Australia); and sodium hydroxide, from Ajax FineChem (Scoresby, VIC, Australia). P4C6 was synthesized in our lab to a purity of >95%, as confirmed by HPLC (Mo et al., 2015 (link)).