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Studies were performed in early passage HUVEC (passage 3-5) and HAo VSMC (passage 4-8) derived from multiple donors.
Hep G2 (human hepatocellular carcinoma), He La (cervical epithelium adenocarcinoma), and JEG-3 (placental choriocarcinoma) cells were obtained from American Type Culture Collection (ATCC, Manassas, VA) and maintained in Dulbecco's modified Eagle's medium containing 10 m—First strand c DNA was synthesized with total cellular RNA (5 μg) derived from varied normal human tissues (Clontech) using random or strand-specific primers and Super Script II reverse transcriptase (Invitrogen).
Recent work has emphasized the importance of post-translational modification of the e NOS holoenzyme (5, 6).
Conversely, treatment of endothelial cells with atheroprotective 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors increases e NOS m RNA levels via transcript stabilization (22, 23).For the further characterization of A and BAC clones were sequenced using an ABI PRISM 377 DNA sequencer (Applied Biosystems, Foster City, CA) and compared with sequences we reported previously (7) and the two assemblies available for human chromosome 7 sequence (27, 28).To characterize were isolated from the RPCI-22 bacterial artificial chromosome library created in the p BACe3.6 vector using female murine 129S6/Sv Ev Tac strain diploid genomic DNA.RNA interference-mediated inhibition of s ONE expression in vascular smooth muscle cells increased e NOS expression.Overexpression of s ONE in endothelial cells blunted e NOS expression.