2D and 3D Viability Assays for Adherent and Spheroid Cells

For 2D adherent cell viability experiment, the cells were seeded at 384-well plate (Corning, no. 3570) at the density of 200 cells/well. The next day, compounds were added using Janus workstation (PerkinElmer). After 5 days treatment, the cell viability was measured by CellTiter-Glo kit (Promega, no. G7570) as the manufacturer recommended. For 3D spheroid assays, NCI-H226 and MSTO-211H cells were plated at the density of 200 cells/well in Ultra-low Attachment (ULA) plate (S-bio, no. MS-9384WZ) without or with 5% Matrigel matrix (Corning, no. 356231) respectively. The cell viability was measured using 3D CellTiter-Glo kit (Promega, no. G9681). The luminescent signal was collected on EnVision plate reader (PerkinElmer). The GR₅₀ values were calculated as previously described Hafner et al., 2016 (link).