2013
DOI: 10.1093/cvr/cvt127
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Shear stress is a positive regulator of thimet oligopeptidase (EC3.4.24.15) in vascular endothelial cells: consequences for MHC1 levels

Abstract: Our findings demonstrate that laminar shear positively regulates endothelial TOP expression. Moreover, a role for ROS production by NADPH oxidase is indicated. Finally, our studies suggest that shear-dependent TOP induction down-regulates MHC1 levels, pointing to a role for TOP in the flow-mediated regulation of endothelial immunogenicity.

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Cited by 13 publications
(15 citation statements)
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“…Quantitation of the final cDNA was determined using a NanoDrop 3300 Spectrophotometer (Thermo Scientific, DE, USA). Amplification of target cDNA sequences using gene-specific primers was subsequently performed and analyzed as previously described [27]. Ribosomal subunit S18 was routinely used for normalization purposes.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Quantitation of the final cDNA was determined using a NanoDrop 3300 Spectrophotometer (Thermo Scientific, DE, USA). Amplification of target cDNA sequences using gene-specific primers was subsequently performed and analyzed as previously described [27]. Ribosomal subunit S18 was routinely used for normalization purposes.…”
Section: Methodsmentioning
confidence: 99%
“…Following experiments, endothelial cell lysates were harvested, resolved by 10% SDS-PAGE under reducing conditions, and electroblotted as previously described [27]. Membranes were blocked for 60 min in tris-buffered saline (TBS: 10 mM Tris pH 8.0, 150 mM NaCl) containing 5% w/v skim milk before being incubated overnight in primary antisera with gentle agitation at 4°C.…”
Section: Methodsmentioning
confidence: 99%
“…Cell lysates were resolved by 10% SDS-PAGE under reducing conditions, and electroblotted as previously described (Rochfort et al, 2014;Guinan et al, 2013). Primary antisera: 0.5 μg/ml anti-ZO-1 mouse monoclonal IgG, 0.5 μg/ml anti-occludin mouse monoclonal IgG, and 0.2 μg/ml anti-GAPDH rabbit monoclonal IgG.…”
Section: Western Immunoblottingmentioning
confidence: 99%
“…Post-treatment, HBMvECs were harvested for total RNA extraction and analysis of ZO-1 mRNA expression as previously described (Guinan et al, 2013) with minor modifications. PCR reaction conditions were: denaturation at 95°C for 10 min followed by 40 cycles of (i) denaturation at 95°C for 15 s, (ii) annealing at 59°C for 60 s, and (iii) elongation at 72°C for 15 s. Each cDNA sample was assayed × 3 and results analysed by the comparative C T method.…”
Section: Quantitative Realtime Pcrmentioning
confidence: 99%
“…Following experiments, endothelial cells were harvested for extraction of total RNA and analysis of TM mRNA expression as previously described (Guinan et al, 2013). Ribosomal subunit S18 was routinely used for normalization purposes.…”
Section: Quantitative Real-time Pcrmentioning
confidence: 99%