Caov3

Ovarian cancer cell lines NIH:OVCAR-3 (OVCAR3) (ATCC^® HTB-161^™), Caov-3 (ATCC^® HTB-75^™) and UWB1.289 (ATCC^® CRL-2945^™) were purchased directly from ATCC and absence of mycoplasma was confirmed independently. All cells were maintained as subconfluent monolayers at 37 °C, 5% CO₂. OVCAR3 cells were grown in high glucose (4500 mg/L) RPMI-1640 (ATCC) with 0.01 mg/mL bovine insulin (MilliporeSigma) and 20% fetal bovine serum (FBS) (MilliporeSigma). Caov-3 cells were grown in modified Dulbecco’s Modified Eagle’s Medium (DMEM) (ATCC) containing 4500 mg/L glucose, 4 mM L-glutamine, 1 mM sodium pyruvate, 1500 mg/L sodium bicarbonate supplemented with 10% FBS. UWB1.289 cells were grown in 1:1 (v:v) mixture of RPMI-1640 (with 2 mM L-glutamine) (Lonza) and mammary epithelial growth medium (MEGM) (PromoCell) and supplemented with 3% FBS. Platinum-resistant UWB1.289 cells (UWB1-PlatR) were established from parental cells by incubating in growth media containing 5 μM carboplatin, in recurrent 48 hour cycles. Following each 48 hour treatment, cells were allowed to recover in platinum-free growth media for 2 to 3 weeks, until typical proliferation resumes, and culture vessel becomes repopulated. A total of 6 cycles were used to establish UWB1-PlatR cells and the stability of the resistant phenotype was verified for a minimum of 8 passages following 6^th treatment cycle.

HGSOC cell lines NIHOVCAR3, CAOV3, and HEY1 were purchased from ATCC. COV318 was purchased from Sigma. OVCAR4 was obtained from the NCI Developmental Therapeutics Program. COV504 cells were a gift from Deborah Marsh (University of Sydney, Australia). Cell line authentication was carried out every 4 years and Mycoplasma testing were performed by-monthly. Cell line passage 1–20 was used. All cells were cultured in RPMI-1640 medium, with 10% FBS and 1% penicillin/streptomycin, at 37°C with 5% CO2. All antibodies used are listed in Supplemental Table 4.