Fig 2: Relative mRNA expression of endocannabinoid signaling-related proteins in the spleen of rats exposed to alcohol during adolescence and control rats grouped by sex.(A) CB1 mRNA levels (fold change); (B) CB2 mRNA levels (fold change); (C) PPARa mRNA levels (fold change); (D) NAPE-PLD mRNA levels (fold change); (E) DGLa mRNA levels (fold change); (F) DGLß mRNA levels (fold change); (G) FAAH mRNA levels (fold change); (H) MGL mRNA levels (fold change); and (I) FAT/CD36 mRNA levels (fold change). The bars represent means ± SEM. The data were analyzed by two-way ANOVA and significant main effects or interaction between factors was indicated as follows: (*) p<0.05 and (**) p<0.01 denote significant main effect of adolescent alcohol exposure; (###) p<0.001 denotes significant interaction between adolescent alcohol exposure and sex. (aaa) p<0.001 denotes significant differences compared to female control rats after post hoc tests; (bbb) p<0.001 denotes significant differences compared to female alcohol-exposed rats after post hoc tests.

Fig 3: Expression of CB1, CB2, PPARa and NAPE-PLD proteins in the spleen of rats exposed to alcohol during adolescence and control rats grouped by sex.Relative protein expression and representative immunoblots of: (A) CB1 protein (fold change); (B) CB2 protein (fold change); (C) PPARa protein (fold change); and (D) NAPE-PLD protein (fold change). Representative immunohistochemical images at 4x magnification of: (E) CB1 protein; (F) CB2 protein; (G) PPARa protein; and (H) NAPE-PLD protein. The bars represent means ± SEM. The data were analyzed by two-way ANOVA and significant main effects or interaction between factors was indicated as follows: (*) p<0.05 denotes significant main effect of adolescent alcohol exposure; (#) p<0.05 denotes significant interaction between adolescent alcohol exposure and sex. (a) p<0.05 and (aa) p<0.01 denote significant differences compared to the respective control rats after post hoc tests; (b) p<0.05 denotes significant differences compared to female alcohol-exposed rats after post hoc tests. The black arrows indicate positive cells. Abbreviations: RP = Red pulp; VS = Venous sinus; WP = White pulp.