# Single molecule fluorescence in-situ hybridization

## Description

Figure 2 Figure 3 Figure 5A-B, E-F Figure S2 Figure S3 Figure S5A Mice: Animals were sacrificed either directly from the homecage (0hr) or 1hr following their first exposure to cocaine (20 mg/kg, IP; ‘Acute’). ‘Repeated 1hr’ mice were sacrificed an hour after their 5th daily exposure to cocaine, compared to ‘Repeated 0hr’, which received 4 days of cocaine, and were sacrificed on the 5th day directly from home cage. ‘Challenge’ mice were sacrificed after 21 days of abstinence from repeated exposure to cocaine, either directly from the homecage (0hr) or 1hr after one final re-exposure to cocaine. n=3 mice in each group. In all experiments 0hr and 1hr are different groups of mice, from the same cohort. Sections: Egr2: Acute (0hr; n=9, 1hr; n=9), Repeated (0hr; n=9, 1hr; n=9), Challenge (0hr; n=6, 1hr; n=8), Fos: Acute (0hr; n=6, 1hr; n=6), Repeated (0hr; n=6, 1hr; n=6), Challenge (0hr; n=4, 1hr; n=6), Drd1: Acute (0hr; n=6, 1hr; n=6), Repeated (0hr; n=6, 1hr; n=6), Challenge (0hr; n=4, 1hr; n=5), Drd2: Acute (0hr; n=6, 1hr; n=6), Repeated (0hr; n=6, 1hr; n=6), Challenge (0hr; n=4, 1hr; n=5). Staining: The smFISH protocol was implemented according to manufacturer guidelines (ACD RNAscope fresh frozen tissue pretreatment and fluorescent multiplex assay manuals; catalog #320513, #320293; as in Gonzales 2019).