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V1.3 patch clamp assay, see: whole cell patch clamp recording was performed with n ≥ 2 individual experiments at each compound concentration using different cells. Four or more different concentrations were determined per dose-response curve
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V1.3 patch clamp assay, see:. Grissmer S., Nguyen A.N., Aiyar J., Hanson D.C., Mather R.J., Gutman G.A., Karmilowicz M.J., Auperin D.D., and Chandy K.G. Mol. Pharmacol. 45 (1994) 1227 whole cell patch clamp recording was performed with n ≥ 2 individual experiments at each compound concentration using different cells. Four or more different concentrations were determined per dose-response curve
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63149086772
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note
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2+ concentration was adjusted to 1 μM. Whole cell patch clamp recording was performed with n ≥ 2 individual experiments at each compound concentration using different cells. Four or more different concentrations were determined per dose-response curve.
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A general PBMC assay setup is described in (a), the readout was performed using BrdU Cell proliferation ELISA kit (Roche) according to manufacturer's instructions
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5 cell/mL and incubated with the compounds or the corresponding amount of vehicle (DMSO) for 30 min. Intracellular calcium increase was induced with 25 μM thapsigargin (Sigma-Aldrich) and was measured using FACSCalibur (BD Bioscience). For the importance of calcium signaling in T-cell proliferation, see:
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5 cell/mL and incubated with the compounds or the corresponding amount of vehicle (DMSO) for 30 min. Intracellular calcium increase was induced with 25 μM thapsigargin (Sigma-Aldrich) and was measured using FACSCalibur (BD Bioscience). For the importance of calcium signaling in T-cell proliferation, see:. Vig M., and Kinet J.-P. Nat. Immunol. 10 (2009) 21
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HaCaT keratinocytes were seeded in KBM/10% FCS and incubated for 24 h at 37 °C. Compounds were diluted in KBM/FCS with a final concn of 1% DMSO, added to the HaCaTs in triplicate and incubated for 48 h. For a readout on cell numbers, the Cell Titer Viability Assay from Promega was used.
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