FIGURE 7.

Treatment with NF-κB inhibitors Bay 11-7082 or MG-132 prevented miR-155 increase in response to alcohol. A, RAW 264.7 macrophages were treated with alcohol for 48 h and treated or not with LPS for 30 min, and nuclear proteins were subjected to EMSA. B and C, for NF-κB inhibition, RAW 264.7 macrophages were pretreated with Bay11-7082 (0.1 μm) or MG-132 (0.25 μm) or DMSO as a negative control for 30 min and then exposed or not to alcohol (50 mm) for 48 h and further treated or not with LPS for 6 h. Expression of miR-155 was assayed by qPCR, and data were normalized to sno202 control. The -fold increase in the expression of miR-155 versus non-stimulated cells is shown. Data represent the mean value (S.E. indicated by error bars) of at least three independent experiments. D, Kupffer cells isolated from pair-fed or alcohol-fed animals (n = 8) were pooled and treated or not with 100 ng/ml LPS for 30 min, and 10 μg of whole cell lysate were subjected to EMSA to evaluate NF-κB activation. E, for NF-κB inhibition, Kupffer cells from pair-fed or alcohol-fed animals (n = 10) were pretreated with MG-132 (2.5 μm) or DMSO for 30 min and further stimulated or not with 100 ng/ml LPS for 6 h. Expression of miR-155 was assayed by qPCR, data were normalized to sno202 control, and the -fold increase in the expression of miR-155 in Kupffer cells from alcohol-fed mice versus Kupffer cells from pair-fed mice is shown. Data represent the mean value (S.E. indicated by error bars). Statistically significant differences are shown (*, p < 0.05).