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. 2017 Nov 27;7:16374. doi: 10.1038/s41598-017-12562-6

Figure 4.

Figure 4

Baicalein inhibited NF-κB pathway in vitro. Cells were treated with dose range of baicalein for 2 h prior to LPS (1 µg/ml) treatment for an additional 24 h. (A) The production of NO in RAW264.7 cells induced by LPS was measured as described in the Methods. (B) Protein level of RAW264.7 cells was determined with antibody against iNOS (1:1000) and β-actin (1:2000 dilution) by immunoblotting. Quantification of the protein expression was performed by densitometric analysis of the blots. Expression was normalized to β-actin. (C) NF-κB promoter-driven luciferase activity in RAW264.7 cells was determined using a luciferase assay system as described in the Methods. Results were expressed as fold values of control cells. (D) mRNA expression of iNOS, COX-2, IL-1α and IL-1β in THP-1 cells was determined by qRT-PCR. Expression was normalized to β-actin. (E) NF-κB p65 nuclear translocation in RAW264.7 cells was evaluated by immunofluorescence staining and images were captured by a fluorescence microscope. Scale bar corresponds to 50 μm and applies throughout. Data were expressed as mean ± SD of three independent experiments (n = 3). ###p < 0.001 vs. vehicle-treated group; **p < 0.01, ***P < 0.001 vs. LPS-treated group.