E plate view 96

BMDMs were plated onto an E-plate view 96 (Acea, Roche), which was placed into a real-time cell analysis (RTCA) SP station (Acea, Roche) in an incubator on 37 °C with 5% CO₂. Subsequent proliferation of BMDMs measured as change in electrical impedance was assessed with xCELLigence RTCA (Acea, Roche) and analyzed with RTCA Software 1.2.

For impedance measurements we used the xCELLigence System (Roche) that provides a highly sensitive method to measure the cell index as a proxy of cell adhesion, proliferation, cell death and morphological alterations. Cell lines were subconfluent, and each cell line has been optimized in previous titration experiments so that cell type-specific optimal conditions for the seeding rate and confluence were obtained for the real-time measurement in these experiments. Cells were seeded at 5000 cells per well in quadruplicates in E-Plate VIEW 96 (Roche) and left untreated for 24 hrs within the RTCA SP station positioned in a 37°C incubator with 5% CO₂ supply. For monitoring compound mediated cytotoxicity, cells were then treated with DMSO only or with 0.2 μM, 1 μM or 5 μM olaparib and incubated for an additional 72 hrs. The Cell Index was plotted and analysed using RTCA software 1.2.1 (Roche).