Fig. (4) Schematic illustration of a multiplexed assay approach that sequentially measures cytotoxicity by a protease release assay and protein aggregates by a laser scanning cytometer plate reader in the same assay plate. From left to right, the uninduced Q103-GFP cells were seeded into an assay plate and incubated for 24 hr (1). The inducer tebufenozide, as well as compounds to be screened, were added to the cells and incubated for 48 hr (2). The protease detection reagent was added to the assay plate to detect the luminescent signal for cytotoxicity resulting from Q103-GFP expression (3). Detergent (final 0.25% Triton X100) was then added to lyse the cells and release soluble proteins into solution. Fluorescent protein aggregates remained attached to the bottom of assay plate were measured by the laser scanning cytometer plate reader (4).