Fig. 6

Acute melatonin reduced the level of activated p-NF-_KB/p-IKKβ in the acute ethanol-treated rat pups and in BV2 cells that were exposed to ethanol. a Western blot analysis of p-NF-_KB65 and p-IKKβ in the rat pups. The bands were quantified using Sigma Gel software, and the differences are presented in a histogram. β-Actin was used as a loading control. n = 10 pups/group, and the number of experiments = 3. b Representative histogram indicates the ELISA analysis of NF-_KBp65 level in the brain homogenates of the rat pups. n = 10 pups/group, and the number of experiments = 3. c Representative image of the p-NF-_KB65 immunofluorescence reactivity in the cortices and CA1 regions of the hippocampi in the rat pups. n = 5 pups/group, and the number of experiments = 3. Magnification × 40, Scale bar = 50 μm. d Western blots and the densitometric analysis of the NF-_KB and IKKβ expression levels in the BV2 cells that were subjected to Nrf2 siRNA and treated with ethanol (100 mM) and melatonin (100 μM) for 12 h. β-actin was used as a loading control. The data are expressed as the mean ± SEM. e A representative histogram indicates the ELISA analysis of NF-_KBp65 level in the BV2 cells that were subjected to Nrf2 siRNA and treated with ethanol (100 mM) and melatonin (100 μM) for 12 h. The number of experiments = 3. The data are expressed as the mean ± SEM. The data are presented relative to control values. Significance = P < 0.05. σ Significantly different from the control saline-treated rat pups; Φ significantly different from the ethanol-treated rat pups. Similarly for in vitro studies, σ significantly different from the non-treated BV2 cells; Φ significantly different from the ethanol-exposed BV2 cells, and ω significantly different from the ethanol+melatonin-exposed BV2 cells