GSM8775930: ScrStat3; Homo sapiens; RNA-Seq
Isolatedbrain endothelial cells were harvested from animals and homogenized in TRIzol solution for RNAseq analysis according to the manufacturer's instructions. Glass slides of HUVECs were removed from the flow chamber and cells were directly lysed in 400µl of TRIzol (Invitrogen, 15596026) per slide and stored in RNase-free microcentrifuge tubes, which were transferred to dry ice for storage until all samples were collected. Total RNA was extracted using the RNeasy mini kit (Qiagen, 74104) according to manufacturer's directions. RNA quality was confirmed using an Agilent Tapestation 4200, and high-quality RNA (RIN > 8.0) was used for RNA-seq library preparation with the Illumina® Stranded mRNA Prep kit (Illumina, San Diego, CA) RNA libraries were multiplexed and sequenced with 100‐bp paired reads (PE100) on an Illumina NovaSeq 6000 platform with a target depth of 30 million reads per sample. Samples were demultiplexed using bcl2fastq conversion software