Centro xs3 microplate luminometer

Huh7.5 cells stably expressing Firefly luciferase (Huh7.5 Fluc) were cultured in Dulbecco’s modified minimum essential medium (DMEM, Life Technologies, Carlsbad, CA, USA) containing 2 mM glutamine, 1× minimum essential medium nonessential amino acids (MEM NEAA, Life Technologies), 100 μg/mL streptomycin, 100 IU/mL penicillin (Life Technologies), 5 μg/mL blasticidin and 10% fetal bovine serum. Cells were maintained in a 37 °C environment with 5% CO₂ supply. Cells were infected with Jc1-derived Renilla reporter viruses in the presence or absence of compounds as described previously [25 (link)]. Infected cells were lysed and then frozen at −80 °C for 1 h following measurements of Renilla and Firefly luciferase activities on a Centro XS3 Microplate luminometer (Berthold Technologies, Bad Wildbad, Germany) as indicators of viral genome replication and cell viability, respectively.

Huh7.5 cells stably expressing Firefly luciferase (Huh7.5 Fluc) were cultured in Dulbecco’s modified minimum essential medium (DMEM, Gibco, Thermo Fisher Scientific, Schwerte, Germany) containing 2 mM L glutamine, 1% minimum essential medium nonessential amino acids (MEM NEAA, Gibco, Thermo Fisher Scientific, Schwerte, Germany), 100 μg/mL streptomycin, 100 IU/mL penicillin (Gibco, Thermo Fisher Scientific, Schwerte, Germany), 5 μg/mL blasticidin and 10% fetal bovine serum. Cells were maintained in a 37 °C environment with 5% CO₂ supply. Cells were infected with Jc1-derived Renilla reporter viruses in the presence or absence of compounds as described [44 (link)]. Infected cells were lysed and then frozen at −80 °C for 1 h following measurements of Renilla and Firefly luciferase activities on a Berthold Technologies Centro XS3 Microplate Luminometer (Bad Wildbad, Germany) as indicators of viral genome replication and cell viability, respectively.

This assay was carried out as described previously by Mulwa et al. [18 (link)]. Huh7.5 cells stably expressing Firefly luciferase (Huh7.5 Fluc) were cultured in Dulbecco’s modified minimum essential medium (DMEM, Life Technologies, Darmstadt, Germany) containing 2 mM L glutamine, 1 × minimum essential medium nonessential amino acids (MEM NEAA, Life Technologies), 100 μg/mL streptomycin, 100 IU/mL penicillin (Life Technologies), 5 μg/mL blasticidin and 10% fetal bovine serum. Cells were maintained in a 37 °C environment with 5% CO₂ supply. Cells were infected with Jc1-derived Renilla reporter viruses in the presence or absence of compounds as described previously [19 (link)]. Infected cells were lysed and then frozen at −80 °C for 1 h following measurements of Renilla and Firefly luciferase activities on a Berthold Technologies Centro XS3 Microplate Luminometer (Bad Wildbad, Germany) as indicators of viral genome replication and cell viability, respectively.