Effects of Developmental Exposure on Neuroimmune Interactions

All data were analyzed with SigmaStat statistical software (Systat Software Inc., San Jose, CA, United States). Three-way (DEP X LPS X Sex) ANOVAs were performed first, and following an interaction with Sex, two-way (DEP X LPS) ANOVAs within each sex were used to assess differences in regional volume (mm³), average cell volume (μm³), the total number of cells/mm³, and the number of cells (of the 2 different morphologies)/mm³. Following interactions, post hoc comparisons (Fisher’s LSD) were performed to further distinguish between groups. For neuron counts and microglial-neuron overlap, two-way (DEP X Sex) ANOVAs were performed to assess differences in the number of neurons, number of microglia-neuron interactions, microglial volume (μm³), and overlap volume (μm³). Interactions were followed up with Fisher’s LSD post hoc tests. The 3D reconstruction data were log-transformed for analysis due to unequal variance, but are displayed as raw data in the figures. Significance was generally assumed at p < 0.05; however, higher-order interactions at p ≤ 0.1 were used to direct the subdivision of data for further analysis (Snedecor and Cochran, 1967 ). These interactions were only reported if subsequent lower-order ANOVAs and/or post hoc tests achieved significance at p < 0.05 (Slotkin and Seidler, 2015 (link)).