GSM7867850: cerebellar endothelial cells - chronic model - Ccm3 knock-out 3; Mus musculus; RNA-Seq
For acute disease model: Endothelial cells were isolated from the cerebella of 3 wild-type and 3 Ccm3 knock-out mice with the Adult Brain Dissociation kit (130-107-677; MiltenyiBiotech), CD45 MicroBeads (30-052-301; MiltenyiBiotech) and CD31 MicroBeads (30-097-418; MiltenyiBiotech) according to the manufacturer's instructions. Total RNA was extracted from each sample with the RNeasy Micro Kit (Qiagen) in the automated QIACube (Qiagen) platform. For chronic disease model: Endothelial cells were similarly isolated from the cerebella of 5 wild-type and 5 Ccm3 knock-out mice. For acute model: RNA sequencing libraries were prepared with SMARTer Stranded Total RNA Sample Prep kit - Mammalian kit (Clontech), according to the manufacturer's instructions. This kit includes depletion of ribosomal RNA prior to library synthesis, barcoding and amplification. 1 μl of each library was analyzed in the Agilent 2100 bioanalyzer for validation of fragment size and quality control of the amplified RNA-seq library, using the High Sensitivity DNA kit (Agilent).  For chronic model: Libraries were prepared with the stranded RNA library prep kits (Collibri; for Illumina), with rRNA depletion kits used according to the manufacturer instructions (#A39003024, A39003096; Invitrogen)