shRNA mediated knock-down of Tal1 in human Endothelial Colony Forming Cells (ECFCs)

Endothelial colony-forming cells (ECFCs) have been reported as promising cells for regenerative medicine thanks to their angiorepair properties. Transcription factors are primary determinants of the functional capacity of the cells and TAL1 has been shown as a critical regulator of endothelial lineage in both development and adult life. However, only few (three) TAL1 targets have been identified so far in mouse and human endothelial cells. This microarray experiment, where TAL1 expression was knocked-down, was designed to identify TAL1-dependent genes in primary human endothelial stem/progenitor cells. ECFCs were isolated from three independent cord blood samples (n=3, biological replicates) and cultured in complete EGM-2 medium. The knockdown of TAL1 was induced by infection with lentiviruses expressing an anti-TAL1 shRNA. A scrambled shRNA was used as a negative control. Cells were then harvested for RNA extraction. DNA-free total RNA was isolated with RNeasy Mini Kit and hybridized to the Affymetrix Human Gene 1.0 ST gene expression microarray.