Fig. 9: Proximity analysis of Th cells within 50 µm of each ILC subtype in thymus (A, C) and spleen (B, D) (n = 8 independent biological samples for each tissue type, 1 mIF staining batch per tissue type).

Heatmaps show the percentage of Th1, Th2 or Th17 (red gradient) according to the ILC1, ILC2 or ILC3 distance (blue gradient, 5 µm steps) in thymus (A) and spleen (B, left panel). Bars show the percentage of Th cells within 50 µm of a single ILC1, ILC2 or ILC3 in the thymus (A) and spleen (B, left panel). In the spleen, where NK/ILC1ie cells were abundant, proximity analysis was also computed for NK/ILC1ie cells versus the three ILC and Th subtypes (B, right panel). C, D Schematic overview of the proximity analysis data. Each circular diagram represents the proximity of each “small” cell (Th cells for thymus and spleen, and ILCs for spleen only) within a 50 µm radius (2 × 25 µm concentric) of the center of the target cell nucleus drawn in the middle (ILC1, ILC2, and ILC3 for thymus and ILC1, ILC3 and NK/ILC1ie for spleen). The mean number and location of each “small” cell are shown schematically within the 2 × 25 µm radius proximity of the target cells (ILC1, ILC2, and ILC3 for thymus and ILC1, ILC3 and NK/ILC1ie for spleen). The small mirrored pie charts represent the percentages of each “small” cell population (Th cells for thymus and spleen, and ILCs for spleen only) within (when located inside the circular diagram, dark grey pie part) or over (when located outside the circular diagram, dark grey pie part) the 50 µm radius of the target cells (ILC1, ILC2, and ILC3 for thymus and ILC1, ILC3 and NK/ILC1ie for spleen). Caption in “C”. Detail of IndicaLabs Halo distribution map with proximity analysis visualization output in thymus. Source data is provided as a Source Data file. Partially created in BioRender. Bezombes, C. (2025) https://BioRender.com/shbs5qf.