Transcriptomic landscape of the adrenal gland of an adult male mouse.

A) Uniform manifold approximation and projection (UMAP) visualisation of adrenal gland spots with manual annotation based on molecular marker prediction for each cluster. BAT – brown adipose tissue, WAT – white adipose tissue, CT – connective tissue, ZG – zona glomerulosa, ZF – zona fasciculata, ZX – X-zone. B) Adrenal glands stained with hematoxylin and eosin (H&E). C) Clusters from the UMAP representation were depicted as color-coded dots on the representative spatial transcriptomic adrenal slide. D) Reconstruction of distinct cell populations within mouse adrenal slices based on UMAP cluster generation. Heat map showing the top 10 expressed (z-score normalised) genes in each obtained cluster. E) Selected representative histological slides with overlaid spots showing expression profiles characterised by the obtained clusters.

Bubble plots of the top enriched GO terms and KEGG pathways for each cluster.

X-axis represents the different clusters corresponding to adrenal gland regions or cell types (ZG, ZF, ZX, medulla, CT, WAT, BAT). Y-axis corresponds to the enriched GO terms and KEGG pathways, grouped by ontology categories: A) Biological Processes (GO BP), B) Molecular Functions (GO MF), C) Cellular Components (GO CC) and D) KEGG pathways. Bubble Size indicates the number of DEGs associated with each term.

Spatial distribution of identified biomarkers for adrenal gland components.

The Cell-type Assignment by Regression on Distances (CARD) analysis based on the list of markers from single-cell data within mouse adrenals. A) The cell proportion matrix mapped onto the adrenal gland spatial transcriptomics analysis. Each cell type is colour-coded, revealing distinct clustering patterns. B) Cell type proportion correlation of selected groups of cells. C) Visualizations of the marker gene expression in ZG, ZF, ZX and Medulla with the spatial locations within the tissue. Fib - fibroblasts, Endoth - endothelial cells, WBC - white blood cells, RBC - red blood cells.

Pseudotime (trajectory) analysis of spots within the adrenal cortex and connective tissue capsule.

A) UMAP dimensionality reduction with visualisation of the spots in the adrenal cortex and connective tissue clusters. B) Trajectory graph generated from the UMAP dimensionality reduction of the spots coloured by pseudotime, according to the colour scale representing the start and end states of the trajectory. The colour gradient from purple to yellow indicates pseudotime progression. C) The range of pseudotime in the analysed clusters (connective tissue, ZG, ZF, ZX) ordered by mean pseudotime value. D) Examples of genes whose expression changes along pseudotime in analysed spots.

Inferred communication networks of individual L-R (ligand-receptor) pairs associated with signalling at A) Wnt, B) non canonical WNT, C) SHH, D) IGF, E) MIF, F) GALECTIN, G) EDN, H) GALANIN and I) NPY pathways.

The left panel shows circle plots illustrating communication strength between or within the analyzed groups, where the thickness of the connecting lines represents the cumulative communication probability. The right panel displays the spatial expression of ligand (left side) and receptor (right side) pairs with the highest communication probability between analysed groups for each specific signalling pathway.