Ebola VLPs containing a beta-lactamase-fused VP40 and GP were prepared in Dr. García-Sastre’s lab, as previously described.6 (link) LiveBLAzer FRET–B/G Loading Kit and CCF2-AM, Dulbecco’s modified Eagle’s medium (DMEM), and Opti-MEM reduced serum medium were purchased from Life Technologies (Carlsbad, CA, USA). An ATP content cell viability assay kit was purchased from Promega (Madison, WI, USA). 1536-well polystyrene plates were purchased from Greiner Bio-One (Monroe, NC, USA). Compounds were purchased from Sigma-Aldrich (St. Louis, MO, USA), Santa Cruz (Dallas, TX, USA), ChemBridge Corporation (San Diego, CA, USA), Enamine Ltd (Kiev, Ukraine), Maybridge Chemical Company (Altrincham, United Kingdom), Vitas-M Laboratory (Champaign, IL, USA), Ambinter (Orléans, France) and AKos Consulting & Solutions Deutschland GmbH (Steinen-Schlächtenhaus, Germany) at the highest available purity. All of the compounds were dissolved as a 10 mM stock solution in dimethyl sulfoxide (DMSO) and diluted in DMSO at a 1∶3 dilution to generate eleven concentrations in 384-well plates, followed by reformatting into one 1536-well compound plate for high throughput screening.