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61449166217
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note
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R = 42 min).
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15
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61449200752
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note
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13C NMR, see Table S1 (Supplementary data).
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25844469333
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0021061819
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note
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2. Cells were harvested and analyzed 9 or 24 h after each treatment by reverse transcriptase-polymerase chain reaction (RT-PCR) or Western blot analysis. The cytotoxic activity of compounds against HaCaT cells was examined by a MTT method (Mosmann, T. J. Immunol. Method 1983, 65, 55).
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19
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61449175217
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note
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11 Statistical analysis. The statistical analyses were performed using the SPSS program (SPSS 12.0). One sample t-test was used to examine the difference between groups. p Values of 0.05 were considered to be statistically significant.
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61449259325
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note
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12,13
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21
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0037087402
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note
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NF-κB reporter gene assay. Cells were plated at in 12-well plate and transfected at 70% confluency. A dual-luciferase reporter assay system (Promega, Madison, WI) was used to determine promoter activity. Briefly, cells were transiently transfected with pNF-κB-luciferase plasmid and phRL-SV plasmid using the Hillymax reagent (Dojindo Molecular Technologies, Gaitherburg, MD). The cells were then incubated in culture medium without serum for 18 h. The firefly and hRenilla luciferase activities in the cell lysates were measured using a luminometer (LB941, Berthold Technologies, Bad Wildbad, Germany). The relative luciferase activity was calculated by normalizing the promoter-driven firefly luciferase activity to the hRenilla luciferase activity (Majumdar, S.; Lamothe, B.; Aggarwal, B. B. J. Immunol. 2002, 168, 2644).
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61449192107
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