Rho kinase paralog regulation of gene transcription

The serine/threonine protein kinase paralogs ROCK1 & 2 have been implicated as essential modulators of angiogenesis; however their paralog-specific roles in endothelial function are unknown. Microarray analysis of ROCK knockdown lines revealed overlapping and unique control of global transcription by the paralogs, and a reduction in the transcriptional regulation of just under 50% of VEGF responsive genes. MS1 mouse endothelial cells were stably transfected with non-targeting, ROCK1 specific, or ROCK2 specific shRNA plasmids. Cells were grown to confluence and treated with sham or 2.5 ng/ml human recombinant vascular endothelial growth factor (VEGF) for 12 hours. RNA from six independent biological replicates from each engineered cell line were collected, pooled, and subjected to triplicate technical replicates of microarray analysis.