GSM4233249: 2131_Scl_Ab_17; Mus musculus; RNA-Seq
RNA sequencing (RNA-seq) analyses were performed on mice tissues in  satellite animals. At scheduled necropsy, a region of the aortic arch matching the scan region for µCT was collected using polymerase chain reaction clean procedures. In addition, the innominate, left carotid, and left subclavian arteries were each sectioned at their junctions with the aortic arch. The lumen of the aorta was flushed with RNase-free saline and the adventitial fat gently scraped away using a sterile cotton swab. The aortic tissue was then washed briefly in RNase-free saline, placed in cryo-tubes, snap-frozen in liquid nitrogen, and stored at –80°C. Samples were subjected to dry pulverization before being homogenized in buffer (350 µL of Qiagen RLT buffer with 10% BME [Qiagen, Germantown, MD, USA]). The homogenate was transferred to an RNase-free 1.5 mL centrifuge tube, after which 95 µL RNase-free water and 5 µL of a 20 mg/mL Proteinase K solution were added. Samples were incubated at 55°C for 10 minutes, centrifuged, and the supernatant collected. RNA was then extracted using an RNeasy Micro Kit (Qiagen) with on-column DNase treatment (Qiagen) according to the manufacturer's instructions. RNA concentration and integrity were assessed using a Bioanalyzer (Agilent, Santa Clara, CA, USA). Samples with ≥80 ng total RNA that had RNA integrity numbers ≥7 were used for sequencing. Library preparation and RNA-seq was performed by Q2 Solutions (Morrisville, NC). Sequencing libraries were created using the IlluminaTruSeq Stranded mRNA method.
GEO Accession
GSM4233249