ATP-Binding Pocket-Targeted Suppression of Src and Syk by Luteolin Contributes to Its Anti-Inflammatory Action

Figure 2

The effects of luteolin on iNOS, COX-2, and TNF-α gene expression and transcriptional regulation in LPS-treated RAW264.7 cells. ((a) and (b)) RAW264.7 cells (5 × 10⁶ cells/mL) were incubated with LPS (1 μg/mL) in the presence or absence of luteolin for 6 h. iNOS, COX-2, and TNF-α mRNA levels were determined using RT-PCR (a) and real-time PCR (b). (c) RAW264.7 cells (5 × 10⁶ cells/mL) were incubated with LPS (1 μg/mL) in the presence or absence of luteolin for the indicated times. After preparing nuclear fractions, the levels of total translocated transcription factors (p65, p50, c-Fos, and c-Jun) were determined by immunoblotting analysis. (d) HEK293 cells cotransfected with NF-κB-Luc (1 μg/mL) and β-gal (as a transfection control) plasmid constructs were treated with luteolin in the presence or absence of adaptor molecule (MyD88) for 12 h. Luciferase activity was determined via luminometry. All data are expressed as the mean ± SD of experiments. and compared to the control group.

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