hiPSC-astrocytes and primary astrocytes were seeded in astrocyte medium 1 day before the experiment. Cells were treated for 24 hr with 50 ng/mL or 100 ng/mL of poly(I:C) (InvivoGen, no. tlrl-pic), 10 μg/mL or 50 μg/mL of LPS (Sigma-Aldrich, no. L5886), and 5 μM or 10 μM human β-amyloid (Aβ42) (CaliforniaPeptide, no. 641-15) and vehicle control solutions (saline for poly(I:C) and LPS or Tris-HCl for Aβ42). Samples were analyzed with IL-6 ELISA assay (Affymetrix eBioscience, no. 88-7066), Human Inflammatory Cytokines Multi-Analyte ELISArray (Qiagen, no. MEH-004A), RT2 Profiler PCR Arrays (Qiagen), and Proteome Profiler Human Cytokine Array (R&D Systems, no. ARY005B). For a detailed protocol see Supplemental Experimental Procedures.
