Size separation of proteins in CM of PDT-treated T-Ep.
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Detection of the FSA activity by chromatographed COS-1 cell-derived IL-1α is included in (B and C).
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Soluble mediator secretion by normal and OA synovial tissue explants (STEs) with or without IL-1α.
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Graphs demonstrate all soluble mediators which were significantly (p<0.01) different between A. normal (black bars) and OA (grey bars) STEs and B. normal and OA STEs under pro-inflammatory (IL-1α) conditions.
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Cell Culture
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VSMCs, EL4, HEK, and HeLa cells were cultured in DMEM and Jurkat and THP-1 cells in RPMI 1640, all supplemented with penicillin, streptomycin, L-glutamine, and 10% FCS.
- Human monocyte-derived macrophages were differentiated as described previously (2, clarified, and stored at −80°C.
- Cells were also made necrotic by incubation with 7-BIO (25 μM) or digitonin (0.1%) (data not shown) or by overnight hypoxic exposure.
- To activate inflammasomes, cells were treated with LPS (1 μg/ml; 4 hr), followed by ATP (5 mM) or Nigericin (20 μM) for 30 min.
- Calpain activity was determined with Calpain-Glo .
- VSMCs, HEK, and THP-1 cells were transfected with pcDNA3 with nucleofection or FugeneHD .
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Bacterial stimuli and TLR ligands induce distinct immune responses in fetal, neonatal and adult keratinocytes.
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IL1α was used as positive control, cell culture medium (untreated) and TSB as negative controls.
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TAK1 exerts its functional effects through activation of NF -κB pathways by phosphorylation at Ser412 (A) Western blot analysis demonstrated that p-TAK1 at Thr184/187 was just found in HEK293 and Hela cells only, while p-TAK1 at Ser412 was generally found in all cell types including ovarian cancer cell lines (n=4).
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After 48 hours, Human IL-1α (10ng/ml) was used to treat the transfected cells with various time points.
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