Author: Mauthe, Mario; Langereis, Martijn; Jung, Jennifer; Zhou, Xingdong; Jones, Alex; Omta, Wienand; Tooze, Sharon A.; Stork, Björn; Paludan, Søren Riis; Ahola, Tero; Egan, Dave; Behrends, Christian; Mokry, Michal; de Haan, Cornelis; van Kuppeveld, Frank; Reggiori, Fulvio
Title: An siRNA screen for ATG protein depletion reveals the extent of the unconventional functions of the autophagy proteome in virus replication Document date: 2016_8_29
ID: iuqa0yrw_51
Snippet: Cells in 96-well plates were washed with PBS and incubated with 50 µl lysis buffer (Thermo Fisher Scientific) at room temperature for 15 min before storing the cell lysates at −20°C. 25-µl aliquots of thawed cell lysate were then used to measure either Firefly or Renilla luciferase expression (depending on the gene carried by the assayed virus) using either the Firefly or Renilla luciferase flash assay kit (both from Thermo Fisher Scientific.....
Document: Cells in 96-well plates were washed with PBS and incubated with 50 µl lysis buffer (Thermo Fisher Scientific) at room temperature for 15 min before storing the cell lysates at −20°C. 25-µl aliquots of thawed cell lysate were then used to measure either Firefly or Renilla luciferase expression (depending on the gene carried by the assayed virus) using either the Firefly or Renilla luciferase flash assay kit (both from Thermo Fisher Scientific). Alternatively, Renilla luciferase activity was measured in the following reaction buffer: 45 mM EDTA, 30 mM sodium pyrophosphate, 1.425 M NaCl, and 10 µM coelenterazine h (Promega; Baker and Boyce, 2014) .
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