Olaparib-Treated Cell Line Characterization

The UWB1.289 (CRL-2945; female) cell line was accessed from internal GSK collections and was obtained from ATCC. The cell line was additionally authenticated using the Promega Cell ID system. The generated short tandem repeat (STR) profiles matched exactly the expected STR profiles of the ATCC lines. Reagents were obtained from Gibco unless stated otherwise. Cells were cultured at 37 °C, 5% CO₂ in 1:1 RPMI:MEGM (Lonza, #CC-3151), 3% fetal bovine serum (FBS, Gibco), MEGM SingleQuots supplements (Lonza, #CC-4136) used without gentamycin-amphotericin. For the Olaparib (CAS: 763113-22-0) treatment, three 15 cm dishes per condition were prepared by seeding 3‒4 × 10⁶ cells per plate; 24 h later medium was removed and 25 ml fresh medium containing dimethylsulfoxide (DMSO, Sigma) or 4uM Olaparib was applied to the cells. Cells were incubated for the indicated time (24 h) at 37 °C, 5% CO₂. Cells were collected by trypsinization, washed twice with PBS, and counted using a Casy Cell Counter (OMNI Life Science). Cell pellets were generated containing 1–2 million cells for either proteomic and phosphoproteomic or transcriptomic analysis. Three independent replicates were generated per data point per readout.