GSM7034227
Pericytes were removed and endothelial cells were lysed directly with Buffer RLT Plus (Qiagen, cat# 1053393) on the inserts. For 1 sample in further analyses, cell lysates from 2 inserts were pooled. RNA was isolated using RNeasy Plus Micro Kit (Qiagen, cat# 74034) containing an integrated gDNA eliminator spin column according to the manufacturer's protocol. For library preparation, samples with 1 µg of purified RNA were used. A total of 8 libraries (4 control and 4 cARLA) were constructed using the Rapid MACE-Seq kit (GenXPro GmbH, Frankfurt, Germany) according to the manufacturer's protocol. Fragmented RNA was reverse transcribed into cDNA, using barcoded oligo(dT) primers containing TrueQuant unique molecular identifiers, followed by template switching. Library amplification was done using polymerase chain reaction (PCR), purified by solid phase reversible immobilization beads (Agencourt AMPure XP, Beckman Coulter, cat# A63882).