Fig. 2

Ethanol increases miR let-7b, HMGB1 in hippocampal-entorhinal slice culture (HEC). HEC slices were exposed to ethanol for 48 h. Tissue was collected for miRNA, mRNA, protein, or immunohistochemistry. Levels of miR let-7b, IL-1β, and TNFα mRNA were measured in brain slice tissue using RT-PCR. TLR7 immunoreactivity (TLR7 + IR) was performed and mean pixel density measured. a TLR7 mRNA expression was increased by ethanol in a dose dependent fashion, up to twofold control levels. b A 23% increase in TLR7 + IR (protein) was found (105.5 ± 1.9 vs 85.3 ± 6.7 mean pixel density ± SEM, Ethanol vs Control, **p = 0.01, t test, N = 5–6 per group). c HEC was exposed to ethanol (25–100 mM) for 48 h. Ethanol increased HMGB1 protein (by ELISA) in the slice tissue by up to 85% in a concentration-dependent fashion. (One-way ANOVA F(4,5) = 48.07, p < 0.0005, Dunnett’s multiple comparisons post-test *p < 0.02,***p < 0.001). d miR let-7 was increased 2.4-fold at 48 h of ethanol exposure (237 ± 16.0% vs 100 ± 2.4%, Ethanol vs Control, mean ± SEM, **p < 0.01, t test