HTS cell viability assay

Cells were seeded at 500 cells/well in 7 μl media in a 1536-well culture plate (Corning, Cat#3893) using a Multidrop Combi Reagent Dispenser (ThermoScientific) with the first column as a medium only control (blk). The next day test compounds (0.1 ul) were dispensed into wells in each plate using a Sciclone ALH 3,000 liquid handler (PerkinElmer) from a compound stock plate to give the indicated final concentrations. Each sample was tested with four replicates. After 3 days of incubation, Cell Titer Blue (1 μl; Promega, G8081) was added to each well using the robotic liquid dispenser. The plate was incubated for 1–4 h at 37 °C. The FI of each well was read using an EnVision Multilabel plate reader (Ex 545 nm, Em 615 nm; PerkinElmer). % of control was calculated using the equation (FI_compound−FI_{Avg. Blk})/(FI_{Avg. Neg}−FI_{Avg. Blk} × 100).