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. 2020 May 6;6(19):eaax7060. doi: 10.1126/sciadv.aax7060

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Fig. 4 — (A) Experimental design. Activated mPFC ensembles were tagged after a last alcohol SA session (Alc SA Tag; N = 5) or after exposure to an NC (NC Tag; N = 5). Animals were perfused 90 min after a cue-induced reinstatement test to investigate colocalization of Fos expression (induced by the test) and hM4Di-mCherry in the NC-tagged and alcohol SA–tagged ensemble. (B) Example images of Fos⁺ and mCherry⁺ cells and colocalization of both markers (white arrowheads). Scale bar, 50 μm. (C) Fos expression in the total neuronal population (Nissl⁺) in the mPFC did not differ between the NC-tagged group (8.2 ± 0.8%) and alcohol SA–tagged group (8.9 ± 1.6%). (D) Percentage of Fos⁺ within the mCherry⁻ and mCherry⁺ population demonstrates enhanced colocalization of Fos and the alcohol SA–tagged ensemble. *P = 0.044. All bar graphs, means + SEM.