GATA2-AS1 siRNA Knockdown in HUVECs

HUVECs were grown to 90% confluency on 60-mm (2 (link)) gelatin-coated tissue culture plates and transfected with siRNA at a final concentration of 40 nM using Oligofectamine (Invitrogen) in a total volume of 2000 μl. Transfection occurred for 4 h at 37 °C in Opti-MEM medium (Invitrogen), after which M199 medium (Invitrogen) containing fetal bovine serum (Hyclone), heparin, and endothelial cell growth supplement (Biomedical Technologies) was added. Cells were incubated with siRNA for 24 to 48 h depending on the downstream application. Custom Stealth siRNAs (Invitrogen) were used to knockdown GATA2-AS1. SiGENOME Non-Targeting siRNA #3 (Dharmacon) was used as control siRNA transfection (Table S3).