*4.2. Cell Cultures*

J774.1 cells were purchased from the Korean Collection for Type Cultures and maintained in DMEM (Gibco BRL, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (Gibco BRL), 100 μg/mL streptomycin, and 100 U/mL penicillin (Gibco BRL). Cells were incubated at 37 ◦C in a humidified 5% CO2 incubator. J774.1 cells were subjected to the following treatments: (i) control; (ii) LPS (Escherichia coli O111:b4) + ATP; (iii) 2.5 μM juglone + LPS + ATP; (iv) 5 μM juglone + LPS + ATP; (v) 10 μM juglone + LPS + ATP. For these treatments, cells were incubated in the presence of juglone for 2 h, followed by treatment with LPS (1 μg/mL) for 6 h and ATP (5 mM) for 1 h. After treatment, cells were harvested for further analyses.
