Author: Michael T. Parker; Smita Gopinath; Corey E. Perez; Melissa M. Linehan; Jason M. Crawford; Akiko Iwasaki; Brett D. Lindenbach
Title: Innate Immune Priming by cGAS as a Preparatory Countermeasure Against RNA Virus Infection Document date: 2018_10_3
ID: j1gkl43g_29
Snippet: To detect cGAMP bioactivity, 2 µL of S1 sample, synthetic cGAMP (positive 548 controls), or DPBS (negative controls) were incubated with 10 6 THP-1 cells, 2 mM 549 ATP, 1.5 ng/µL SLO (a kind gift from Dr. N. Andrews, University of Maryland) and 550 media in 8 µL (total volume). After 1.5 hours at 30°C, reactions were lysed with an 551 equal volume of RIPA buffer and processed for phosphorylated IRF3 western blot, 552 as above. 553.....
Document: To detect cGAMP bioactivity, 2 µL of S1 sample, synthetic cGAMP (positive 548 controls), or DPBS (negative controls) were incubated with 10 6 THP-1 cells, 2 mM 549 ATP, 1.5 ng/µL SLO (a kind gift from Dr. N. Andrews, University of Maryland) and 550 media in 8 µL (total volume). After 1.5 hours at 30°C, reactions were lysed with an 551 equal volume of RIPA buffer and processed for phosphorylated IRF3 western blot, 552 as above. 553
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