Migration assays
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HUVECs were seeded at 15–20×103/well and cultured to 90% confluence on 24-well transwell tissue culture inserts (24-well plate with inserts, 5 µm pore/6.5 mm, TC-treated) as previously described 3 receptor agonists and antagonists were present during the migration in both transwell compartments.
- Migration experiments displayed an inter-donor variability in the absolute number of migrating T cells, thus for each experiment migration data have been normalized considering the number of counted cells in control conditions as 100%.
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Islet isolation, culture, and exposure to different beta-cell inhibitory substances
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Following the initial culture period, islets were cultured for an additional 6–24 hours in CMRL 1066 containing antibiotics, 2 mM glutamine and one of the following supplements: 0.5 mM sodium palmitate solubilized in 0.5% (weight/volume) fatty acid and lipopolysaccaride free bovine serum albumin (BSA) ; recombinant human Interleukin 1beta (IL-1ß (50 units/ml) and Interferon-gamma (IFN-γ) (1,000 units/ml) ; 100 mM hydrogen peroxide; 2 mM DETA/NO or 10 mM streptozotocin (STZ) .
- To some of the groups 10 µM of Imatinib was added at different time points prior to the addition of test substances given above.
- To controls equal amounts of vehicle (DMSO) were supplemented.
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The placenta releases sST2.
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C) Placental explants release sST2, secretion was increased by treatment with TNFα and IL-1β (100 ng/ml) and hypoxia-reperfusion injury, and was decreased under hypoxic conditions (n = 3).
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Effect of CORM-2 on HO-1 protein (A) and mRNA expression (B) in OA synoviocytes.
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Cells were stimulated with IL-1β (10 ng/ml) for 24 h and 16 h in the presence or absence of CORM-2 (50, 100, 200 µM) or RuCl3 (200 µM).
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Functional characterization of mutant N1.
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Cells were treated with IL-1β, lysed and the relative fold activation of NF-κB activity was determined.
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