Fig. 2.

Placental cytokine expression and factor analysis across gestational age, genotype and MIA conditions. A Heatmaps of log-transformed concentrations of 18 cytokines/chemokines in placental tissue, stratified by genotype (XXF, XXM, XYF, XYM), gestational age (E13.5 early; E18.5 late), and treatment (saline vs. MIA). Blue denotes low expression; red denotes high expression. B Exploratory factor analysis model showing loadings of each cytokine onto five latent factors. Color scale reflects loading strength and direction (–0.3 [indigo] to 0.9 [yellow]). C- G Factor scores for placental cytokine Factors 1–5. Three-way ANOVA revealed main effects and interactions of MIA timing, sex chromosome complement, and gonadal sex. XXF placentas showed elevated Factor 1 (treatment × sex chromosomes × gonad interaction, F(3,109) = 3.12, p = 0.029) and 2 scores (treatment × sex chromosome × gonadal sex interactions, F(3,109) = 5.902, p < 0.001) that persisted with MIA exposure. XYM placentas exhibited suppressed Factor 1–3 scores, particularly following late MIA. Factor 4 scores were higher in gonadal females relative to gonadal males (gonadal sex effect, F(1,109) = 6.847, p = 0.010). No significant effects were observed for Factor 5. Main treatment effects are represented by “t”. 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