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.Targeting the Rho/ROCK axis by the small interfering RNA technique in rat liver sinusoidal endothelial cells.
The Rho-associated kinase (ROCK) family, consisting of ROCK1 and ROCK2, has been identified as a pivotal regulator of the actin-myosin cytoskeleton. ROCK-interacting proteins are crucial to the modulation of ROCK activity. The ROCK inhibitor fasudil has been known to have marked anti-inflammatory activity. However, the intracellular signaling mechanisms of fasudil in sinusoidal endothelial cells (SECs) remain unknown. In the present study, the potency and specificity of small interfering RNA (siRNA) interference against ROCK1 and ROCK2 were examined by fluorescence-activated cell sorting (FACS), immunoblot, and FACS analysis of cellular migration. After siRNA transfection of rat liver SECs, we observed morphological changes that included cytoskeletal reorganization and actin stress fiber formation. Furthermore, the ROCK1- and ROCK2-siRNAs significantly inhibited collagen contraction and adhesion of SECs and stimulated the migratory activity of the SECs. These results suggest that transfection with siRNAs against ROCK1 and ROCK2 may be a promising approach to inhibit the ROCK signaling pathway and allow subsequent investigation of the mechanism by which fasudil works in SECs.Australians are taking a “pay or be sacked” approach to the unemployment rate, with many now expecting to be fired if they don’t get a job.
Key points: Almost 400,000 more people were unemployed in the three months to June, and there are currently one million job seekers
Almost 400,000 more people were unemployed in the three months to June, and there are currently one million job seekers Year-on-year, the unemployment rate increased 0.6 per