Fig. 9
The interaction of MCP-1/CCR2 signaling, TLR4 and GSK3β in response to ethanol exposure. a SIM-A9 cells were pretreated with RS504393 (100 μM) or Bindarit (300 μM) for 12 h, and then exposed to EtOH (0.4%) for 12 h. The expression of GSK3β, phosphorylated GSK3β at serine 9 (phospho-GSK3βS9), phosphorylated GSK3β at tyrosine 216 (phospho-GSK3βY216), and TLR4 were determined by IB and quantified by the normalization to the expression of actin. b C57BL6 mice of PD4 were treated with Bindarit, RS504393, and EtOH as described in Fig. 2. 8 h after EtOH treatment, the expression of TLR4 and its downstream adaptors TRIF and MyD88, and phospho-GSK3βS9 in the brain was determined by IB and quantified by the normalization to the expression of actin. c SIM-A9 cells were pretreated with TAK242 (1 μM) for 12 h, and then exposed to EtOH (0.4%) for 12 h. The expression of GSK3β, phospho-GSK3βS9, phospho-GSK3βY216, MCP-1, and CCR2 was by IB and quantified by the normalization to the expression of actin. d SIM-A9 cells were pretreated SB-216763 (10 μM) for 12 h, and then exposed to EtOH (0.4%) for 12 h. The expression of TLR4, MCP-1, and CCR2 was by IB and quantified by the normalization to the expression of actin. Each data point was the mean ± SEM of three independent experiments. *p < 0.05, statistically significant difference from control group; #p < 0.05, statistically significant difference from EtOH-treated group