Cell viability assays via luminescence

Luciferase-expressing human GBM cells (LN229) stably expressing TAZ^4SA were seeded in 96-well flat-bottom plates at a density of 3000 cells/well alone or cocultured with neutrophils in a 1-to-20 ratio in duplicates and incubated at 37 °C for 15 h. At the end of coculture, luminescence was measured as a readout for cell viability using the Luciferase Assay System (E1500, Promega) following the manufacturer’s instructions. Briefly, cells were washed once with DPBS, lysed, and given a D-firefly luciferin potassium salt dissolved in culture medium prior to luminescence measurement via a multi-mode plate reader (BMG Labtech). Luminescence from tumor cell monoculture was used to establish a baseline reading. All readings were averaged between duplicate wells. Percent survival was calculated by normalizing blank-subtracted luminescence of coculture wells to that of tumor cell monoculture. For all compound treatment experiments, tumor cell monoculture treated with the same compound at the same concentration was used as control for normalization (100% survival).