Exp. 1 Genome-wide transcriptional profiling of CeA during incubation of Meth craving

As shown in Fig. 1a, we performed intravenous surgeries on two groups of rats (total n = 26) and trained them to self-administer either saline (n = 12) or Meth (n = 14) as described in Supplementary Online Material in 2 independent runs. We performed live decapitation on withdrawal days 2 and 35, and collected CeA tissue (as in Fig. 1c) for mRNA preparation. We used the extracted mRNA for library preparation and RNA-seq. We pooled tissue from two rats as one biological replicate. The number of biological replicates in each group was: Day 2: Saline = 3, Meth = 4; Day 35: Saline = 3, Meth = 3.

Tissue collection for subsequent RNA-seq analysis. a Experimental timeline. b Meth and saline self-administration training. Data are mean ± SEM number of Meth (0.1 mg/kg/infusion) infusions and active and inactive lever presses during the ten 9-h daily self-administration sessions for Exp. 1 (total n = 26). During training, active lever presses were reinforced on an FR1 20-s timeout reinforcement schedule, and Meth or saline infusions were paired with a 5-s tone-light cue. c Diagram for orbitofrontal cortex (OFC) and central amygdala (CeA) tissue collection