Fig. 3.

Fetal brain cytokine expression and factor analysis across gestational age, genotype and MIA conditions. A Heatmaps of log-transformed cytokine/chemokine concentrations in fetal brain tissue, stratified by genotype (XXF, XXM, XYF, XYM), gestational age (E13.5 early; E18.5 late), and treatment (saline vs. MIA). Blue indicates low expression; red indicates high expression. B Exploratory factor analysis of fetal brain cytokines identifying three latent factors. Color intensity reflects loading magnitude (0.4 [indigo] to 0.9 [yellow]) for each cytokine’s contribution to Factors 1–3. C-E Bar plots of Factors 1–3 across genotype and treatment groups at early and late gestational timepoints. Across all factors, XYF fetal brains showed elevated Factor 1 (three-way interaction, F(3,109) = 3.383, p = 0.021), Factor 2 (three-way interaction, F(3, 109) = 3.825, p = 0.012) and Factor 3 scores (gonadal sex × sex chromosome interactions, F(1,109) = 8.585, p = 0.004), particularly following late MIA. Main effects of treatment and sex chromosomes indicated “t” and “c”, respectively. Three-way ANOVA with Tukey’s post hoc tests, n = 6–8/group. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. Heatmaps and graphs created with BioRender or GraphPad Prism