RNA-sequencing of Endothelial Cell Lines Expressing GNAQ WT or Mutant Q209L

To investigate the transcriptional effects of GNAQ p.Q209L mutation on endothelial cells, bulk RNA-sequencing was performed on endothelial cells lines expressing GNAQ wildtype or mutant Q209L. Cells were also treated with Trametinib or vehicle control to elucidate the specific effects of MAPK/ERK signaling on these cells. An endothelial cell line (endothelial colony forming cells) was transduced with doxycycline-inducible GNAQ-WT or GNAQ-Q209L constructs. For these experiments, cells were grown in low-serum media (2% FBS in EGM2). To induce gene expression prior to RNA collection, cells were treated with 1µg/mL doxycycline for 18 hours, followed by an additional 6 hours of DMSO (vehicle control) or 10nM Trametinib treatment. Four conditions were analyzed: 1) WT (no dox) – vehicle; 2) WT (dox) – vehicle; 3) Q209L (dox) – vehicle; 4) Q209L (dox) – Trametinib. For each condition, we performed four biological replicates.