Fig. 3

Activation of microglia 24 h after ethanol treatment in the brain of transgenic and wild-type mice. a Relative expression of Aif1 in the brain of hHSPB1-overexpressing and wild-type mice 24 h after ethanol treatment. Relative expression was correlated with the wild-type control group (100%). n = 8 mice per group. b Morphological changes of microglia 24 h after EtOH treatment on sagittal brain sections of hHSPB1-overexpressing and wild-type mice. Microglia with different morphologies were visualized with IBA1 immunostaining using the peroxidase method. Scale bar: 100 μm (OCx and Hip). c Proportions of IBA1+ microglia in different brain regions according to their morphology. d Quantification of IBA1 immunoreactive areas. Results are given in percentage of the immunopositive areas compared to the outlined brain areas (relative area). n = 3 mice per group, 3 sections per animal. e Relative expression of M1 microglia markers (Cd68, iNos) and M2 microglia markers (Mrc1, Arg1) in the brain of hHSPB1-overexpressing and wild-type mice 24 h after ethanol treatment. Relative expression was correlated with the wild-type control group (100%). n = 8 mice per group. Values are presented as mean ± SEM. Asterisk indicates wild-type EtOH group vs wild-type control group; plus sign indicates transgenic EtOH group vs transgenic control group; hashtag indicates transgenic EtOH group vs wild-type EtOH group; white triangle indicates transgenic control vs wild-type control group. Statistical analysis: 2-way ANOVA followed by Tukey post hoc test. *^/#/+/Δp < 0.05; **^/##/++p < 0.01; ***^/###/+++p < 0.001. Ctx cortex, Hip hippocampus, RSCx retrosplenial cortex, OCx occipital cortex, PCx parietal cortex, FCx frontal cortex, Th thalamus, Str striatum