Atp content cell viability assay kit

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.

Ebola VLPs containing a beta-lactamase-fused VP40 protein (EBOV BlaVP40) and GP were produced in Dr. García-Sastre’s lab, as previously described (Tscherne et al., 2010 (link)). LiveBLAzer FRET–B/G Loading Kit with CCF2-AM were purchased from Life Technologies (Carlsbad, CA, USA); eGFP-EBOV was produced as previously described (Johansen et al., 2013 ). Constructs of RFP organelle marker were purchased from Thermo Fisher Scientific. An ATP content cell viability assay kit was purchased from Promega (Madison, WI, USA). Polystyrene plates (384-well and 1536-well black, clear bottom, sterile, tissue culture treated) were purchased from Greiner Bio-One (Monroe, NC, USA). The compounds were purchased from Sigma (St. Louis, MO, USA) at the highest available purity.