Statistical analyses were performed using GraphPad Prism version 10.0 (GraphPad Software, La Jolla, CA). Data are presented as the means ± SEM, unless stated otherwise. Statistical analysis was performed on data derived from biologically independent experimental replicates, as specified in the figure legends. The impact of chronic ethanol application was assessed by normalizing the results to the baseline activity in high-PRS microglia, with changes in high-PRS and low-PRS microglia relative to baseline values defined as relative changes. Differences in the effects of various ethanol concentrations in high-PRS and low-PRS microglia were evaluated using a two-way analysis of variance (ANOVA), followed by post hoc analysis with Fisher’s least significant difference test for multiple comparisons. Within the same group of microglia, the effects of ethanol before and after application were compared using unpaired t tests. Statistical significance was indicated by the following symbols: (*P < 0.05, **P < 0.01, and ***P < 0.001), with (NS) denoting no statistically significant difference.
