Fig. 2

Effects of ethanol on the levels of the different proteins in the WT and TLR4-KO astrocyte-derived EVs. Immunoblot analysis and quantification of the following proteins: a TLR4, IL-1R, COX-2, NFκB-p65; b NLRP3, procaspase-1, and caspase-1 of the EVs secreted by the untreated and ethanol-treated WT and TLR4-KO astrocytes. CD63 was used as a loading control. The WT and TLR4-KO astroglial cells were treated with ethanol (40 mM) for 24 h. A representative immunoblot for each protein and their molecular weight are shown. 40 μg/μL of protein were loaded in each lane. Data represent mean ± SEM, n = 6 independent experiments. *p < 0.05 and **p < 0.01 compared to their respective control group, according to the two-way ANOVA followed by Bonferroni’s post-hoc test. For the TLR4 statistical analysis, *p < 0.05 compared with the control value, according to the t test