Monocytes or RAW 264.7 macrophages were stimulated with Escherichia coli-derived LPS (100 ng/ml), 25 mM ethanol, and the combination of LPS and ethanol at the times indicated in the figure legends. The 25 mM in vitro ethanol concentration approximates a 0.1g/dl blood alcohol level, which is achieved in vivo after a dose of moderate drink and is a little above the legal limit of blood alcohol concentration. Cell viability was not affected by ethanol or LPS treatment. Monocytes and/or macrophages were plated in 6-well plates and exposed to 25 mM alcohol over an extended period of up to 7 days. For short-term or acute alcohol exposure, cells were exposed to 25 mM alcohol for 24 h or less followed by LPS stimulation.
