Fig. 1: Behaviour epoch-specific, brain-wide astrocyte Fos ensemble mapping.

a, Fos⁺ astrocytes were tagged with Cre-dependent mNG AAV in four cohorts (NoFC, FC, NoFR and FR), followed by STPT, 3D reconstruction, segmentation and single-cell counting. i.p., intraperitoneal injection. Scale bar, 1 mm. b, Freezing of shocked (FC, FR and NoFR: n = 12) and non-shocked (NoFC: n = 6) mice (left), and recall freezing shocked (FC and FR: n = 8) and non-shocked (NoFC: n = 6 mice) (right). c, 3D rendering of whole brain, the hippocampus (HPC), amygdala and primary visual cortex (VISp) with Fos-mNG astrocytes. A, anterior; D, dorsal; L, lateral; M medial; P. posterior; V, ventral. Scale bars, 500 µm. d,e, Heat map of fold changes in BAE count (versus NoFC (d) and FR versus FC (e); NoFC: n = 6; FC: n = 4; NoFR: n = 4; FR: n = 4 mice). Abbreviations for brain regions are listed in Extended Data Fig. 6. *False discovery rate (FDR) < 0.05. Levene’s test followed by one-way analysis of variance (ANOVA) or Kruskal–Wallis; post hoc Tukey or Dunn test with Bonferroni correction. FDR controlled by Benjamini–Hochberg procedure. Full data in Supplementary Table 2. f–h, FC- and FR-induced Fos counts in neurons versus astrocytes across engram areas for FC versus NoFC (f), FR versus NoFC (g) and FR versus NoFC with log scales, showing correlation Pearson’s correlation coefficient (r) (h). h, Amygdalar subregions are indicated with yellow dots. BMA, basomedial amygdala; MA, medial amygdala. i, Schematic of AAV tagging of Fos⁺ neurons and BAEs. j, Images of tagged Fos⁺ neurons and BAEs (representative of 10–22 sections from 3–5 mice per group). Scale bar, 200 μm. k, Quantification of Fos⁺ neurons (NoFC: n = 11 slices from 4 mice; FC: n = 10 slices from 3 mice; NoFR: n = 12 slices from 4 mice; FR: n = 13 slices from 4 mice) and BAE density (NoFC: n = 22 slices from 4 mice; FC: n = 18 slices from 3 mice; NoFR: n = 17 slices from 3 mice; FR: n = 15 slices from 5 mice). One-way ANOVA, Tukey’s test. Data are mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; NS, not significantly different. Sample sizes and replicates in Supplementary Table 1.