Extended Data Fig. 2: Aorta transcriptomes for atherosclerosis-associated axon neogenesis in ATLOs.

a-d, Gene expression in WT and Apoe^−/− aorta during ageing. a, Heatmaps of differentially regulated NS development genes (left) and respective NS-related GO terms (right). b-d, Quantification of selected candidate genes for axon neogenesis (b), axon guidance (c), and synaptic transmission (d). n = 3 WT, 3 Apoe^−/− aorta per group. e-h, Gene expression in aged aorta laminae. e, Heatmaps show differentially expressed NS-related GO terms in laser capture microdissection-derived aged WT vs Apoe^−/− aorta adventitia and plaque. f-h, Quantification of candidate genes associated with axon neogenesis (f), axon guidance (g), and synaptic transmission (h). n = 3 WT adventitia, 4 Apoe^−/− adventitia no plaque, 4 ATLO, 3 plaque. i, Schematic view of laser capture microdissection-derived aged aorta laminae, and numbers of statistically significant (t-test with Benjamini-Hochberg correction) and differentially regulated NS development genes in two-tissue comparisons. n = 3 WT, 4 Apoe^−/− no plaque, 4 ATLO, 3 plaque. j, Heatmaps show differentially expressed NS-related GO terms in aged WT and Apoe^−/− RLN vs ATLO. k, Quantification of selected candidate genes. l,m, Comparison of selected candidate NS-genes in aged WT vs Apoe^−/− RLN (l) and spleen (m). n = 3 WT RLN or spleen, 3 Apoe^−/− RLN or spleen, and 4 ATLOs. Cluster analyses were performed using ANOVA with Benjamini-Hochberg correction (a,e,j). Signal intensities and statistics are reported in supplementary Tables 1–5. n, Detection and enumeration of ALDH1⁺ axons (arrow) in WT adventitia vs ATLO (n 3WT, 5 Apoe^−/−). Scale bar, 20 µm. o, Detection of NGF-expressing periaxonal cells (arrow) and non-neuronal cells (arrowhead), and their quantification in WT adventitita versus ATLOs (n = 3 WT, 3 Apoe^−/−). Scale bar, 20 µm. Inset: higher magnification of the boxed area. Scale bar, 8 µm. Data represent means ± s.e.m. n represents biologically independent animals. Generalized linear model with Bonferroni post hoc test (b-d,f-h,k); two-sided unpaired Student´s t-test (n,o); multiple unpaired t-test corrected for multiple comparisons (Bonferroni) (l,m)

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