EMH-induced adrenal glands have a modified stromal architecture with PDGFRα clusters hosting hematopoietic foci with rare c-Kit⁺ HSPCs. (A) Selection of genes enriched in the PDGFRα-expressing cells of the murine adrenal gland (PDGFRα-expressing clusters: mC6 “mesenchymal” in red and mC13 “capsule” in purple, as defined by Bedoya-Reina et al¹⁹). (B) Adrenal plastic-adherent cells are supportive of CD45⁺ cell expansion. Expansion of total hematopoietic cells (CD45⁺) upon coculture for 7 days of fluorescence-activated cell sorted murine HSPCs (LKS) with a confluent feeder layer of BM stromal cells (BMSCs) obtained from flushed, collagenase-digested bones or adrenal gland, in the absence of additional cytokines. Data are represented as mean ± SD and groups were compared with 1-way ANOVA followed by Holms-Šidák multiple correction test (n = 5 for LKS monoculture, 6 for BMSCs, and 11 for adrenal plastic-adherent cells. Data were obtained in 2 independent experiments). (C) Representative images of control and EMH–induced adrenal glands (n = 2-3 mice per group) stained for CD45 (green), PDGFRα (orange) and DAPI (white). Scale bars represent 20 μm in all instances in panel C. (D) Representative images of control and EMH-induced adrenal glands (n = 2, multiple nodules per adrenal) stained for LEPR (blue), PDGFRα (orange), and endomucin (teal). Yellow arrowheads indicate examples of colocalization of PDGFRα and LEPR (colocalization in pink), with the latter having an elongated pattern marking cells in pericyte position, whereas white arrowheads indicate adrenal parenchyma cells that display a different pattern of LEPR, mostly perinuclear. Scale bars represent 20 μm in all instances in panel D, except for the inset, in which it represents 5 μm. (E) Representative images of control and EMH-induced adrenal glands (n = 3, multiple nodules per adrenal) stained for PDGFRα (orange), cKit (blue), and DAPI (white). Scale bars represent 20 μm in the controls, and 10 μm in the EMH-induced mice. The scale bar in the inset represents 5 μm.