Journal: bioRxiv

Article Title: A highly sensitive cell-based luciferase assay for high-throughput automated screening of SARS-CoV-2 nsp5/3CLpro inhibitors

doi: 10.1101/2021.12.18.473303

Figure Lengend Snippet: A. The structure of the four secondary hit compounds (#5, #24, #35 and #39) is shown and their tertiary amide component is circled. GC376 was used as a control. B. The ability of compounds #5, #24, #35 and #39 to inhibit SARS-CoV-2 replication was evaluated at concentrations corresponding to 3-fold serial-dilutions from 50 to 0.02 μM in a multicycle replication assay on A549-ACE2 cells, followed by RT-qPCR quantification of the viral genomic RNA in the supernatant collected at 72 hpi. The percentages of viral inhibition was determined as described in the section. (one representative experiment of two independent experiments is shown). C. In parallel the indicated compounds were tested on A549-ACE2 cells for cell cytotoxicity by the CellTiter-Glo assay. In B and C, the data are expressed as the mean ± SD of technical triplicates (one representative experiment of two independent experiments is shown).

Article Snippet: HEK-293T cells (purchased from ATCC, # CRL-11268) and A549 cells stably expressing the human ACE2 receptor (A549-ACE2, kindly provided by O. Schwartz, Institut Pasteur) were grown in complete Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% fetal bovine serum (FBS).

Techniques: Quantitative RT-PCR, Inhibition, Glo Assay