Fig. 1: Context-exposed withdrawal increases cocaine-associated seeking.

A Schematic depicting experimental design; cocaine withdrawal/extinction (WD/Ext) modalities followed by transcriptomics (RNA-seq) in NAc-subregions. B, C Acquisition of cocaine SA and Test 1 (T1; drug-free). D, E Separation of the experimental groups undergoing WD/Ext in the previous drug context (Ctx), in the home-cage (HC), or under full extinction conditions (Ext), followed by Test 2 (T2; drug-free). Repeated measures and two-way ANOVAs, followed by Sidak post-hoc; **p < 0.0001, **p < 0.0001, respectively. F, G Comparison between Tests 1 and 2 displayed increased (ns), unchanged, and reduced seeking behaviors in the Ctx, HC and Ext groups, respectively. These effects were observed in active and inactive lever pressing (total lever pressing). Repeated measures, followed by Sidak post-hoc; **p < 0.0001, **p = 0.005, **p < 0.0001 and ^#p = 0.10, **p < 0.0001. H–J Percent change analysis from Tests 1 to 2 confirms that Ctx-exposed rats trended to display incubation-like behavior, while the HC and Ext groups remained stable or reduced their seeking, respectively (one-way ANOVA, followed by Sidak post-hoc; **p < 0.0001). This interpretation was supported by the percentage (%) distribution of rats within the experimental groups (Fisher exact test, **p = 0.003, **p’s < 0.0001 (vs. Ext)). Cocaine, Ctx: n = 12; HC: n = 17; Ext: n = 18. All data are shown as mean ± SEM. **p ≤ 0.01, *p ≤ 0.05, ^#p ≤ 0.10, ns=non-significant. Created in BioRender. Holt, L. (2025) https://BioRender.com/l78p336 (HY27SN20NE); Prism 9: GPS-1216108-L###-#####.