94). The cell counts obtained for each ROI in the different sections of each animal were averaged to calculate the mean number of c-Fos-positive cells within a particular brain region of that animal. These average
values/brain region of each animal were used for statistical analysis. Statistical evaluation of the results DAPT molecular weight was made with SPSS 20 (SPSS Inc., Chicago, Illinois, USA). In general, the data were analyzed by one-way or two-way analysis of variance (ANOVA), as appropriate, in some cases for repeated measurements. Two-way ANOVA was performed with the NOD agonists (VEH, MDP, FK565) and LPS (VEH, LPS) as the between subject variables in order to reveal significant main factor effects or interactions denoted as NOD × LPS interactions. The homogeneity of variances was assessed with the Levene test. In case of sphericity violations the Greenhouse–Geisser correction was applied. Post-ANOVA analysis of group differences was performed with the Tukey HSD (honestly significant difference) test, when the variances were homogeneous, and with the Games–Howell test, when the variances were unequal. In
case of a non-parametric distribution of the parameters, statistical differences among groups were determined with the Kruskal–Wallis test and post-hoc analysis of group differences was performed with the Mann–Whitney test. p values were adjusted for multiple comparisons with the Bonferroni correction. Probability values of p < 0.05 were regarded as statistically significant and p < 0.1 were regarded Enzalutamide clinical trial as a trend. All data are presented as means + SEM, n referring to the number of mice in each group. MDP, FK565 and LPS altered locomotion, exploration, food intake and SP in a compound-, combination- and time-dependent manner (Fig. 2). Repeated measures ANOVA revealed a significant interaction of NOD (VEH, MDP, FK565) × LPS (VEH, LPS) × time (days post-treatment) for the variation in locomotion
(F(5.661,116.05) = 2.457, p < 0.05). The same was true for exploratory behavior (F(5.250,110.25) = 2.470, p < 0.05). Likewise, there was a significant NOD × LPS × time interaction for the differences in food intake (F(5.025,105.52) = 5.244, p < 0.001). SP depended on time (F(1.130,39.55) = 27.838, p < 0.001), with a significant interaction pheromone with LPS (F(1.130,39.55) = 18.397, p < 0.001) and an interaction with the NOD agonists by trend (F(2.260,39.55) = 2.339, p = 0.10). Post-hoc analysis revealed significant NOD × LPS interactions on day 1 and 2 post-treatment. While MDP (1 mg/kg) and FK565 (0.001 mg/kg) alone did not induce any significant changes in locomotion, LPS (0.1 mg/kg) led to a decrease of locomotion for 2 days after injection when compared with the VEH-treated group. Combination of MDP + LPS attenuated locomotion compared to treatment with MDP or LPS alone during day 1 and 2 post-treatment (Fig. 2A). Likewise, the combination of FK565 + LPS significantly decreased locomotion when compared with FK565 or LPS alone.