Illumina HiSeq 4000 paired end sequencing; RNA-Seq of human primary microglial cells treated with vehicle or human paired helical filaments (PHFs) purified from human tauopathy autopsy brain
Human microglial cells (ScienCell, Cat. #: 1900) were thawed and plated in a  6-well tissue culture plate pre-coated with 0.1mg/ml of Poly-L-Lysine (Sigma, P4707). The culture media was changed 16 hours after thawing and  cells remained undisturbed for another 24hours.  Human microglial cells were treated with vehicle or 2ug/ml of hPHF (paired helical filaments isolated from human AD brain) in triplicates for 24 hours. RNA was extracted from cells using Qiagen Rneasy mini kit with DNase treatment as per manufacturer's instructions.  Library construction was performed by Quick Biology Inc. (Pasadena, CA) using KAPA Stranded RNA-Seq Kits with RiboErase.