For competition cell culture assays, Cas9-expressing cells were transduced with the ipUSEPR sgRNA (RFP-positive) or ECEG cDNA (GFP-positive) constructs in 96-well plates at ~50% infection. The cell cycle was measured by Click-iT™ Plus EdU Alexa Fluor 647 Assay Kits (Invitrogen). The cellular apoptosis was detected using Annexin V-FITC Apoptosis Detection Kit (Invitrogen) and Active Caspase3 Staining Kit (Invitrogen). The cell viability was detected by 4’,6-diamidino-2-phenylindole (DAPI; Invitrogen) dye exclusion. The δ score of drug combination was defined as:
where V%(0), V%(T), V%(K), and V%(T+K) are the observed cell viability under no drug, tunicamycin alone, KDS alone, and tunicamycin plus KDS combination, respectively. Data were obtained by high-throughput flow cytometry using an Attune NxT flow cytometer with an autosampler (ThermoFisher).
where V%(0), V%(T), V%(K), and V%(T+K) are the observed cell viability under no drug, tunicamycin alone, KDS alone, and tunicamycin plus KDS combination, respectively. Data were obtained by high-throughput flow cytometry using an Attune NxT flow cytometer with an autosampler (ThermoFisher).