Viability and apoptosis assays

For viability assays, ~10,000 cells were plated per well into 96-well plates. Drugs were added for the incubated period before replacing with fresh medium. One day later, viability was measured using the CellTiter 96 Aqueous One Solution Cell Proliferation Assay (Promega) according to the manufacturer’s instructions. For Annexin V/PI flow cytometry analysis of apoptosis, drugs were added to ~400,000 cells per well in 6-well plates for indicated time points. Cells were harvested and stained with the Dead Cell Apoptosis Kit (ThermoFisher Scientific) according to the manufacturer’s instructions. Samples were processed on a BD FACSCanto II and results were analyzed using Kaluza. Caspase 3/7 activity was measured using the Caspase-Glo 3/7 assay system (Promega) according to the manufacturer’s instructions. Drugs were added to ~10,000 cells per well in 96-well plates for 4 h after which caspase-glo reagent was added. Caspase activity was measured 1 h later.