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Cocultures of hASMCs and CFBE (1:5 ratio) were transfected with a FITC-conjugated siRNA (see Materials and Methods).
Human aortic vascular smooth muscle cells (HASMCs) were obtained from the National Infrastructure of Cell Line Resource (Beijing) and cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco) which contained 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, at 37[degrees]C in a humidified atmosphere with 5% C[O.sub.2].
Effect of Plasma from AD Patients on Apoptosis of HASMCs. The mRNA levels of Bcl2, Bax, and [alpha]-SMA in these three groups were measured; the results showed that the Bax mRNA levels were significantly increased in the Ang II group and further increased when treated with plasma from AD patients; the opposite trend of mRNA levels of Bcl2 and a-SMA was observed (Figure 4).
Human aortic smooth muscle cells (HASMCs) (Cell Applications, USA) were cultured in DMEM/F12 (50:50) and supplemented with L-glutamine, antibiotic/antimycotic cocktail, and 10% fetal bovine serum (FBS).
HASMCs were seeded in 24-well plates and allowed to grow until they reached 30-40% confluency.
HASMCs were seeded in 96-well plates and allowed to grow until they reached 30-40% confluency.
Our results demonstrated that fluid shear stress decreased the proliferation rate of hASMC [ILLUSTRATION FOR FIGURE 1 OMITTED].
The effects of flow on nitrite production by hASMC are shown on Figure 2.
This result provided further evidence that NOS II plays no role in flow-induced nitrite production by hASMC. Monoclonal or polyclonal antibodies against NOS II showed no immunoreactivity with Western blot analysis [ILLUSTRATION FOR FIGURE 3 OMITTED], which verified that the inducible isoform was not present, either in control or sheared hASMC cultures.
HASMCs (Cascade Biologics, OR, USA) were grown and passaged as described previously [23, 24].
Western blot analysis was used to determine the changes in cell levels of HO-1 in HASMCs. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membrane, and then the membranes were blocked with 5% milk.
This inhibits TNF-[alpha]-induced MMP-9 expression and the migration of human aortic smooth muscle cells (HASMCs) .
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