From COVID-19 to the common cold: Scientists identify broadly-effective, infection-halting compound ubcnews
) matched T-1686568-induced viral titer reduction. Our collective findings across multiple assays strongly demonstrate GSK3β inhibition may serve as an effective antiviral strategy for early intervention in COVID-19 and other coronavirus infections, and recommend a potent GSK3β inhibitor for further translational studies.Caco-2 cells , Calu-3 cells and Vero E6 cells were cultivated in accordance with ATCC recommendations. Human hepatoma Huh-7.5.
] and passaged in Vero E6 cells. For experiments, passage 3 of the virus was used with a viral titer of 1.5 × 10plaque forming units /mL. B.1.617.2 and BA.1 variants obtained from BEI Resources and passaged in Vero E6 cells. HCoV-229E was kindly obtained from Dr. Eric Jan, and infections were caried out in a Biological Contamination Level 2+ laboratory in Huh-7.5.1 cells at 33 °C. Cells were seeded at a concentration of 10,000 cells/well in 96-well plates, 24 h prior to infection.
All of the protein kinases used in this study were active preparations of recombinant, GST-fusion human proteins expressed inor Sf9 insect cells and sourced from SignalChem .
Peptide affinity-purified rabbit polyclonal antibodies directed against synthetic peptides were acquired from Kinexus Bioinformatics and based on the following SARS-CoV-2 proteins: nucleocapsid aa 156-170 , ORF1a aa 735-750 – NSP2 , and spike aa 574-588 .All protein kinase reactions were performed with the ADP-Glo Kinase Assay from ProMega . Substrate peptides were assayed at a final concentration of ~ 250 μM with 250 μM ATP for 30 min at 30 °C in a final volume of 25 μL.
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