Fig. 2: Behavioral phenotype of adolescent offspring exposed to prenatal stress and stratification into vulnerable and resilient subgroups.

Pregnant dams were exposed to prenatal stress (PNS) or left undisturbed (CT). At adolescence (from PND35 to PND39) the resulting offspring were subjected to the following behavioral test: the social interaction test, displayed as percentage of social preference (panel A), the sucrose preference test, displayed as percentage of sucrose preference (panel B), and the novelty suppressed feeding test, displayed as latency time to eat the food (panel C). A two-step cluster analysis including the three behavioral tests (the latency to eat the food in the NSF, the social preference in the SI, and the sucrose preference in the SP test) from control (CT n = 25) and prenatally stressed (PNS n = 46) offspring was performed to identify subgroups of animals with different behavioral outcomes. The pie charts show the cluster distribution (with the percentages and the number of animals in each cluster) for all offspring combined, and for CT and PNS groups separately (panel D). Panel (E) displays the predictor of importance for cluster separation, with NSF having the highest predictor importance. Panel (F) shows the behavioral readouts for CT and PNS vulnerable (PNS-vul) and resilient (PNS-res) offspring of both sexes. Statistical analysis for panels A, B, and C: PNS effect, *p < 0.05, **p < 0.01, ****p < 0.0001, two-way ANOVA; n = 14 to 30 animals per group. Statistical analysis for panel (F): **p < 0.01 vs. the respective CT group and ^####p < 0.0001, statistically different from the respective CT and PNS-res groups (one-way ANOVA, Tukey’s post hoc); n = 5 to 19 animals per group. The data are presented as the mean ± SEM.