Author: Alisha Chitrakar; Sneha Rath; Jesse Donovan; Kaitlin Demarest; Yize Li; Raghavendra Rao Sridhar; Susan R. Weiss; Sergei V. Kotenko; Ned S. Wingreen; Alexei Korennykh
Title: Realtime 2-5A kinetics suggests interferons ß and ? evade global arrest of translation by RNase L Document date: 2018_11_26
ID: mxdvdw9u_23
Snippet: HeLa cells in 10 cm plates at 80% confluency were treated with 1 μg/mL poly-IC for 6 hours and lysed with 1 ml RIPA buffer (Thermo Fisher Scientific) supplemented with 0.5 mM PMSF protease inhibitor. The lysate was spun down at 12,000 g for 10 minutes at 4°C. The supernatant was passed through a 3 kDa centrifugal filter to collect the fraction of small nucleic acids. Equivalent amounts of small RNA from mock and . CC-BY 4.0 International licens.....
Document: HeLa cells in 10 cm plates at 80% confluency were treated with 1 μg/mL poly-IC for 6 hours and lysed with 1 ml RIPA buffer (Thermo Fisher Scientific) supplemented with 0.5 mM PMSF protease inhibitor. The lysate was spun down at 12,000 g for 10 minutes at 4°C. The supernatant was passed through a 3 kDa centrifugal filter to collect the fraction of small nucleic acids. Equivalent amounts of small RNA from mock and . CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/476341 doi: bioRxiv preprint poly-IC treated samples were used in the luminescence assays. 2-5A activity was tested using 32 p-5'-radiolabeled RNA substrate and recombinant human RNase L as described previously 22 .
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