Cytotoxicity Evaluation of Compounds

To determine the cytotoxicity of the compounds, 70 to 80% confluent 293T cells in a 96-well plate were incubated with serial dilutions of each compound (up to 100 μM) for 5 h. Cell cytotoxicity was determined by a CytoTox 96 nonradioactive assay kit (Promega, Madison, WI) by measuring a cytosolic enzyme lactate dehydrogenase, which is released upon cell lysis, following the manufacturer’s instructions, and the 50% cytotoxic concentration (CC₅₀) value for each compound was calculated using GraphPad Prism software. The nonspecific cytotoxic effects of these compounds were also reported previously (30 (link), 31 (link), 40 (link), 43 (link), 44 (link)).

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Perera K.D., Johnson D., Lovell S., Groutas W.C., Chang K.O, & Kim Y. (2022). Potent Protease Inhibitors of Highly Pathogenic Lagoviruses: Rabbit Hemorrhagic Disease Virus and European Brown Hare Syndrome Virus. Microbiology Spectrum, 10(4), e00142-22.

Publication 2022

CytoTox 96 nonradioactive assay kit

293t cells Assay Cell Cytosolic Cytotoxicity Dilutions Enzyme Lactate dehydrogenase Prism Promega

Corresponding Organization : Kansas State University

Other organizations : University of Kansas, Wichita State University

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Variable analysis

independent variables

Serial dilutions of each compound (up to 100 μM)

dependent variables

Cell cytotoxicity determined by a CytoTox 96 nonradioactive assay kit (Promega, Madison, WI) by measuring a cytosolic enzyme lactate dehydrogenase, which is released upon cell lysis
50% cytotoxic concentration (CC50) value for each compound

control variables

293T cells in a 96-well plate were 70 to 80% confluent
Compounds were incubated with cells for 5 h

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