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. 2014 Mar 17;35(4):496–503. doi: 10.1038/aps.2013.191

Figure 2.

Figure 2

Daidzein decreases TNF-α -dependent Cxcl2 expression. MLE-12 cells were transiently transfected with the reporter plasmids (A) Cxcl2-luc or (B) NF-κB-luc and Renilla-luc, as described in Materials and methods. The cells were cultured and stimulated with TNF-α in the presence or absence of daidzein (10 μmol/L) or PJ34 (2.5 μmol/L) for 6 h. Cxcl2-driven Luc expression and NF-κB transcriptional activation was measured by normalizing the firefly luciferase activity to that of renilla luciferase. The expression in untreated cells was considered 1. (C) MLE-12 cells were exposed or not exposed to TNF-α (in the presence or absence of daidzein or PJ34) for 1 h, RNA was extracted, and real-time PCR was performed to analyze Cxcl2 mRNA levels. (D) MLE-12 cells were exposed to TNF-α for 1 h in the presence of different concentrations of daidzein, as indicated, RNA was extracted, and real-time PCR was performed to analyze Cxcl2 mRNA levels. Data (mean±SD) are representative of three independent experiments (cP<0.01 compared with normal cells; fP<0.01 compared with TNF-α-stimulated cells).