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. 2024 Sep 18;15:8176. doi: 10.1038/s41467-024-52429-9

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Illustration of Nurr1-flx mouse line construct and the viral strategy of Nurr1 deletion in CLA. b ISH autoradiographs (left) and graphs (right) showing Nurr1 mRNA at the CLAs of AAV-Ctrl and AAV-Nurr1cKO mice. (AAV-Ctrl: n = 6, AAV-Nurr1cKO: n = 6; *p < 0.0001, Two-sided Unpaired t-test). c Immunofluorescence showing GFP, Cre, and Nurr1 expression in the CLAs of AAV-Ctrl (upper) and AAV-Nurr1cKO (lower) mice (scale-bar: 200 μm). d Illustration showing the level and the brain structures of the coronal sections that were hybridized on ST arrays. e H&E-stained brain sections (left) and on-tissue visualization of the 25 molecular clusters in AAV-Ctrl and AAV-Nurr1cKO brains (AAV-Ctrl: n = 3, AAV-Nurr1cKO: n = 3). f 2D-UMAP with a categorical color code to visualize the spots’ distribution in the 25 identified molecular clusters. g On-tissue (left) and 2D-UMAP (right) visualization of Nurr1 expression in AAV-Ctrl and AAV-Nurr1cKO sections. h On-tissue (left) and 2D-UMAP (right) visualization of the spots located in the CLA region from AAV-Ctrl and AAV-Nurr1cKO sections. i On-tissue (left) and 2D-UMAP (right) visualization of clusters 8 and 11 in CLA region from AAV-Ctrl and AAV-Nurr1cKO sections. j Schematic depiction of the deconvolution analysis using the dataset from Zeisel et al. 2018. k, l On-tissue (left) and 2D-UMAP (right) visualization of TEGLU12 (k) and TEGLU20 (l) from AAV-Ctrl and AAV-Nurr1cKO sections. Data in line-graphs are expressed as mean ± SEM. Boxplots show all data points, the 25th and 75th percentile (box), the median (center), and the maxima (whiskers). The dashed circle denotes the CLA region. AAV: adeno-associated virus, ISH: in situ hybridization, GFP: green fluorescent protein, OD: optic density, arb: arbitrary units, AI: agranular insula, CLA: claustrum/dorsal endopiriform cortex complex, CP: caudoputamen, AUC: area under the curve, FC: fold change, ST: spatial transcriptomics, CTX: cerebral cortex, CNU: cerebral nuclei, H&E: hematoxylin & eosin staining, dlCP: dorsolateral caudoputamen, dmCP: dorsomedial caudoputamen, ACB: nucleus accumbens, wm: white matter, OT: olfactory tubercle, MSC: medial septal complex, LSX: lateral septal complex, Ep: ependymal cells, ChINs: striatal cholinergic interneurons, TEGLU: telencephalon excitatory projecting neurons. Source data are provided as a Source Data file.