Author: Lamborn, Ian T.; Jing, Huie; Zhang, Yu; Drutman, Scott B.; Abbott, Jordan K.; Munir, Shirin; Bade, Sangeeta; Murdock, Heardley M.; Santos, Celia P.; Brock, Linda G.; Masutani, Evan; Fordjour, Emmanuel Y.; McElwee, Joshua J.; Hughes, Jason D.; Nichols, Dave P.; Belkadi, Aziz; Oler, Andrew J.; Happel, Corinne S.; Matthews, Helen F.; Abel, Laurent; Collins, Peter L.; Subbarao, Kanta; Gelfand, Erwin W.; Ciancanelli, Michael J.; Casanova, Jean-Laurent; Su, Helen C.
Title: Recurrent rhinovirus infections in a child with inherited MDA5 deficiency Document date: 2017_7_3
ID: vipx6t7e_58
Snippet: A549 cells, seeded at 50,000 per well in 24-well tissue culture plates 20 h prior, were transduced with lentivirus stocks for 48 h to similar transduction efficiencies (Dittmann et al., 2015) . In brief, the cell culture medium was replaced with lentivirus particles diluted in F-12K medium (Gibco) containing 8 µg/ml polybrene (Sigma-Aldrich) and spininfected at 1,350 g for 30 min at 35°C. Transduced cells were infected with HRV-B14 at MOI of 1 .....
Document: A549 cells, seeded at 50,000 per well in 24-well tissue culture plates 20 h prior, were transduced with lentivirus stocks for 48 h to similar transduction efficiencies (Dittmann et al., 2015) . In brief, the cell culture medium was replaced with lentivirus particles diluted in F-12K medium (Gibco) containing 8 µg/ml polybrene (Sigma-Aldrich) and spininfected at 1,350 g for 30 min at 35°C. Transduced cells were infected with HRV-B14 at MOI of 1 for 72 h, as described in the HRV infections section. Transduction efficiencies were assessed by flow cytometry after gating on dead negativetransduced (Zombie Aqua; BioLegend; mCherry + ) cells. HRV-B14-infected cultures were washed with DPBS containing calcium and magnesium (Lonza), and pan-HRV qRT-PCR was performed as described in the Pan-HRV and type I/III IFN qRT-PCR section.
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