Supplementary Figure 1: Progression of port entry and freezing responses across training.
From: Amygdala inputs to prefrontal cortex guide behavior amid conflicting cues of reward and punishment
Related to Figure 1. (a-b) Training timeline and behavioral apparatus. Sensory modalities for the sucrose- and shock-predictive cues (conditioned stimuli, CSs) were counterbalanced across animals. During competition trials, the CS-Suc and CS-Shock associations were co-presented to induce conflicting motivational drives and competition between the reward- and fear-related behaviors. (c) Progression of port entry responses during reward conditioning, plotted in blocks of five trials (repeated measures two-way ANOVA with Bonferroni post-hoc tests, epoch x trial-block interaction, F14,448 = 5.36, P < 0.001; t14 > 4.16, ***P < 0.001). (d) Progression of port entry and freezing responses during the discrimination and competition sessions, plotted as session-blocks (trial-type x training-session interactions; port entry during discrimination, F2,60 = 21.4, P < 0.001; freezing during discrimination, F2,60 = 23.4, P < 0.001; port entry during competition, F1,30 = 1.68, P = 0.20; freezing during competition, F1,30 = 7.37, P = 0.011; Bonferroni post-hoc tests, t14 > 3.89, ***P < 0.001). (e-g) Effects of cue modality on behavioral performance. Superimposed dots represent individual subjects. No significant statistical differences were detected between animals that were trained with the light cue for the reward association and the tone cue for the fear association (RLight FTone: n = 8 animals), or vice versa (RTone FLight: n = 8 animals) (Reward Conditioning: two-way repeated measures ANOVA, group x training-session interaction, F2,28 = 0.57, P = 0.57) (Discrimination Session: Bonferroni-corrected unpaired T-Tests, all t14 < 1.71, all P > 0.11) (Competition Session: Bonferroni-corrected unpaired T-Tests, all t14 < 1.61, all P > 0.13). (h) Purity of behavioral responses across trial types during the last competition session. Dots represent individual animals. Values closer to “1” indicate that animals tended to solely perform a behavioral response type (either port entry or freezing) during each trial (see inset heatmap on the left). Values away from “1” indicate that animals tended to perform behavioral transitions (from freezing to port entry, or vice versa) within single trials (see inset heatmap on the right). (i) Effects of previous trial on behavioral output during competition trials. Connecting lines represent individual subjects. No significant differences were detected (repeated measures one-way ANOVA: port entries, F2,15 = 1.81, P = 0.18; freezing, F2,15 = 1.56, P = 0.23). (j) Latency of port entry and freezing responses during competition. Superimposed dots represent latencies for individual subjects. Latencies were capped at 20 s, which was the maximum trial length. Significant differences were detected across trial types for port entry latencies (repeated measures one-way ANOVA with Bonferroni post-hoc tests, F2,47 = 99.7, P < 0.001; all t15 > 5.89, all ***P < 0.001) and freezing latencies (F2,47 = 64.6, P < 0.001; all t15 > 4.29, all ***P < 0.001). In all data panels, error bars represent s.e.m.
