(A) Eugenol antagonized the effects of the activators of autophagy, ERK, p38 MAPK and IKK/NF-κB pathways determined by Western blot. A549 cells were seeded in 6-well plate for 24 h, then treated with or without eugenol (5 µg/mL) and the activators (autophagy activator 5 µM rapamycin, IKK/NF-κB activator 10 µM LPS, ERK activator 100 ng/ml EGF and p38/JNK activator 10 µM Anisomycin), after 24 h, the cells were collected and subjected to western blot using their corresponding antibodies. (B) Eugenol antagonized the promotions of the activators on the dissociation of Beclin1-Bcl2 heterodimer determined by co-IP assay. A549 cells were seeded in 6-well plate for 24 h, and then treated with or without eugenol (5 µg/mL) and the activators (100 ng/ml EGF, 10 µM Anisomycin and 100 µM H2O2), after 24 h, the cells were collected and subjected to co-IP assay. Data shown were the mean ± SD of three independent experiments. *P<0.05 and **P<0.01.