Additional file 5 of Transcriptomics and translatomics identify a robust inflammatory gene signature in brain endothelial cells after ischemic stroke

Additional file 5: Figure S3. Validation of expression of cell-type marker genes by RT-PCR. In independent groups of naïve mice (n = 3 per group) we obtained endothelial mRNA by the RiboTag technique using PdgfbicreER:Rpl22HA mice, or by CD31+ cell sorting, as before. We extracted mRNA and carried out RT-PCR for validation of expression of cell markers. We compared the results with mRNA extracted from whole brain tissue (cortex) of naïve mice (n = 4). Values are expressed as fold vs. total brain tissue. Results show that the two methods of endothelial RNA extraction are enriched in endothelial markers, such as CD31 (Pecam1) and Vegfc. However, the platelet derived growth factor receptor beta, Pdgfrb, a marker of pericytes, is also enriched, and Tubb3, a marker of neurons is enriched in the mRNA obtained from the PdgfbicreER:Rpl22HA mice, confirming some contamination with RNA from other cell types in each preparation. ***p