HEK293T cells were transfected with plasmids harbouring a luciferase gene under an NF-κB promoter (NF-κB-Luc) or AP-1 promoter (AP-1-Luc). To activate the luciferase genes, MyD88 or TRIF genes were co-transfected. A beta-galactosidase (β-gal) reporter gene was also co-transfected as a control for normalization of transfection efficiency. Transfections were performed using the polyethylenimine (PEI) method as reported previously (Hossen et al. 2016; Han et al. 2018). The transfected cells were stabilized for 24 h and then treated with Sb-EE for another 24 h. Luciferase activity was measured with a luciferase assay system as described previously (Woo et al. 2005).

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