Author: Korbinian Bösl; Aleksandr Ianevski; Thoa T. Than; Petter I. Andersen; Suvi Kuivanen; Mona Teppor; Eva Zusinaite; Uga Dumpis; Astra Vitkauskiene; Rebecca J. Cox; Hannimari Kallio-Kokko; Anders Bergqvist; Tanel Tenson; Valentyn Oksenych; Magnar Bjørås; Marit W. Anthonsen; David Shum; Mari Kaarbø; Olli Vapalahti; Marc P. Windisch; Giulio Superti-Furga; Berend Snijder; Denis Kainov; Richard K. Kandasamy
Title: Critical Nodes of Virus–Host Interaction Revealed Through an Integrated Network Analysis Document date: 2019_2_13
ID: ig6ul3u7_12
Snippet: Drug re-purposing screen For the HMPV NL/1/00 screen, approximately 4×10 4 human retinal pigment epithelial (RPE) cells were seeded per well in 96-well plates. The cells were grown for 24 h in DMEM-F12 medium supplemented with 10% FBS, 0.35% NaHCO 3 , and 100 µg/ml streptomicine and 100 IU/ml penicillin. The medium was replaced with virus growth medium (VGM) containing 0.2% bovine serum albumin (BSA), 2 mM L-glutamine, 0.35% NaHCO 3 , and 1 µg.....
Document: Drug re-purposing screen For the HMPV NL/1/00 screen, approximately 4×10 4 human retinal pigment epithelial (RPE) cells were seeded per well in 96-well plates. The cells were grown for 24 h in DMEM-F12 medium supplemented with 10% FBS, 0.35% NaHCO 3 , and 100 µg/ml streptomicine and 100 IU/ml penicillin. The medium was replaced with virus growth medium (VGM) containing 0.2% bovine serum albumin (BSA), 2 mM L-glutamine, 0.35% NaHCO 3 , and 1 µg/mL L-1tosylamido-2-phenylethyl chloromethyl ketone-trypsin in DMEM-F12. HCV screen-associated cell culture conditions are described in . The compounds were added to the cells in 3-fold dilutions at seven different concentrations starting from 50 µM. No compounds were added to the control wells. The cells were mock-or virus-infected at a multiplicity of infection (MOI) of one. After 48 h of infection, the medium was removed from the cells. To monitor cell viability, CellTiter-Glo reagent containing Firefly luciferase and luciferin was added (30 µL per well). This assay quantifies ATP, an indicator of metabolically active living cells. The luminescence was measured with a plate reader. To determine compound efficacy, HMPV NL/1/00-induced GFP expression was measured. The half-maximal cytotoxic concentration (CC 50 ) and the half-maximal effective concentration (EC 50 ) for each compound were calculated after non-linear regression analysis with a variable slope using GraphPad Prism software version 7.0a. The relative effectiveness of the drug was quantified as the selectivity index (SI = CC 50 EC 50 ). Cytotoxicity and antiviral activity of the compounds against GFP-expressing HCV in Huh-7.5 cells was determined as previously described ). preprint 4 author/funder. All rights reserved. No reuse allowed without permission.
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