Cytokine and chemokine responses of PBMCs, SkMCs, HMEC-1, and HFFs were measured using methods previously reported by us19 (link),36 (link),37 (link) with the following Milliplex kits: HCYTMAG-60K-PX41, HCYPMAG-63K, and HCYP2MAG-62K (EMD Millipore, Billerica MA). Cells were seeded at a density of 105 cells per well. Cells were either stimulated for 16 h with graded concentrations of TLR ligands, or mock-stimulated with vehicle (DMSO). Supernatants were collected by Precision 2000 liquid handlers (BioTek, Winooski, VT) and diluted 1:3 for each kit. The data was acquired on a MagPix® instrument and the data analyzed using Milliplex Analyst (EMD Millipore, Billerica MA).
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Cytokine and Chemokine Profiling of Cells
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Corresponding Organization : University of Minnesota
Other organizations : University of Kansas
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Variable analysis
independent variables
- Graded concentrations of TLR ligands
- Vehicle (DMSO) for mock-stimulation
- Cytokine and chemokine responses of PBMCs, SkMCs, HMEC-1, and HFFs
- Cell density (10^5 cells per well)
- Stimulation duration (16 h)
- Positive control: Not explicitly mentioned
- Negative control: Mock-stimulation with vehicle (DMSO)
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