Fig. 3

MCP-1 antagonist RS504393 prevents β-endorphin neuronal apoptosis induced by ethanol-activated microglial exosomes in cultures. β-endorphin neuronal apoptosis was determined by measuring the nucleosome unit (A) or by measuring mean fluorescence intensity (B, C) of β-endorphin neurons in primary cultures following incubation with exosomes obtained from control culture (Exo_Control) or with Exo_Control + RS50493, with 50 mM ethanol treatment (Exo_Ethanol), or with Exo_Control + RS50493. RS504393 (10µM/ml) was applied 3 h prior addition of exosomes. Representative images are shown, β-endorphin neuron are stained Green. Nucleus are stained with DAPI (blue). Scale bar represents 100 μm. Histograms represent mean ± SEM values of mean fluorescence intensity for β-endorphin neurons and are analyzed using using one-way ANOVA followed by Newman-Keuls post hoc test. Significant differences between the groups are marked with *, p < 0.05; **, p < 0.01; ****, p < 0.0001