Research Article

IRAK1/4-Targeted Anti-Inflammatory Action of Caffeic Acid

Figure 3

Effect of CA on transcription factor activation. ((a), (b), (c), and (d) left panel) HEK293 cells that had been cotransfected with NF-κB-Luc, AP-1-Luc, or CREB-Luc plasmid constructs (1 μg/mL each) and β-gal (transfection control) were treated with CA (0 to 400 μM) or CA-containing plant extracts (Na-ME, Vc-ME, and Ep-ME (200 to 400 μg/mL)) in the presence or absence of PMA (100 nM) or forskolin (2 μM). Luciferase activity was measured with a luminometer. (d) HEK293 cell viability was determined by an MTT assay. (e) Levels of p65/NF-κB and AP-1 family proteins (c-Jun, c-Fos, FRA-1, and p-FRA-1) in the nuclear fractions of LPS-treated RAW264.7 cells cultured in the presence or absence of CA (100 to 400 μM) were determined by immunoblotting analyses with antibodies against the total proteins. Relative intensity was calculated using total levels by the DNR Bioimaging system. and , compared to the control.
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