title: RNA-Seq: Aeroplysinin-1 on gene expression in primary human lung endothelial cells with and without influenza A
library_construction_protocol: Total RNA was isolated from human primary microvascular endothelial cells (HPMECs) seeded on 6 well plates (3.8 x 105cells/well). HPMECs were infected with influenza and treated with DMSO, Ap (1 μM) or Ap-1 (a modified inactive derivative of Ap) for two hours.  Lysates were prepared by washing cells twice with PBS+, lysed in recommended volume of RLT Lysis buffer from the Qiagen RNeasy RNA Isolation Kit (Cat. No. / ID: 74104) and collected by scraping with a sterile cell scraper. Lysates were vortexed for 20 seconds, centrifuged at max speed for 3 minutes at room temperature at stored at -80C for up to two weeks prior to extraction with the kit which was performed according to manufacturer's instructions.  Library prep was performed using the NEB Ultra II Directional mRNA kit.