Human astrocytes were grown in culture as described in de Rivero Vaccari et al. in 2012
[10 (link)]. Primary human astrocytes (Lonza Basel, Switzerland) were grown in culture in complete Astrocyte Growth Medium (Lonza Basel, Switzerland) for seven days. RIG-1 signaling was stimulated with 5′ triphosphate double-stranded RNA (5′ppp dsRNA, Invivogen San Diego, CA, USA) as a specific ligand to stimulate RIG-1 signaling at different concentrations (2 and 4 μg/ml) for 18 hours. After stimulation, cells were harvested and immunoblotted for RIG-1 (Anaspec Fremont, CA, USA), phosphorylated IRF3 (Novus Biologicals Littleton, CO, USA), amyloid precursor protein (Abcam Cambridge, MA, USA) and amyloid-β (Epitomics Burlingame, CA, USA) expression as described.
[10 (link)]. Primary human astrocytes (Lonza Basel, Switzerland) were grown in culture in complete Astrocyte Growth Medium (Lonza Basel, Switzerland) for seven days. RIG-1 signaling was stimulated with 5′ triphosphate double-stranded RNA (5′ppp dsRNA, Invivogen San Diego, CA, USA) as a specific ligand to stimulate RIG-1 signaling at different concentrations (2 and 4 μg/ml) for 18 hours. After stimulation, cells were harvested and immunoblotted for RIG-1 (Anaspec Fremont, CA, USA), phosphorylated IRF3 (Novus Biologicals Littleton, CO, USA), amyloid precursor protein (Abcam Cambridge, MA, USA) and amyloid-β (Epitomics Burlingame, CA, USA) expression as described.
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