Extended Data Fig. 4: Quantification of non-epithelial stromal cells subsets and aaTEC gating.

a–c, Concatenated flow cytometry plots and quantities for cell populations within the fibroblast (a), endothelial (b), and “other” (c) cell lineages in 2-mo (n = 10) and 18-mo (n = 10) mice. c, Concatenated flow cytometry plots and quantities for pericytes (PC), vascular smooth muscle cells (vSMC) and mesothelial cells (MEC) (n = 10/age). d, Frequency and numbers of DN-TEC across lifespan: 2-mo (n = 14), 6-mo (n = 5), 9mo (n = 15), 12-mo (n = 5), and 18+mo (n = 18). e, Violin plots with extensive list of aaTEC1 and aaTEC2 markers. f, Gating strategy for aaTECs. aaTEC1 were first gated on CD45⁻TER119⁻ then PDGFRα^-CD31⁻ cells. EpCAM⁺MHCII⁺ cells were gated as the whole TEC compartment, then mTECs and cTECs were excluded by taking the UEA1⁻Ly51⁻ double negative fraction and gating on CLDN3. aaTEC2 were also first gated on CD45⁻TER119⁻ then PDGFRα^-CD31⁻ cells. EpCAM⁻MHCII⁺ cells were then gated and PDPN⁺PDGFRβ^- were classed at aaTEC2. Summary data represents mean ± SEM; each dot represents an individual biological replicate. Statistics for a–c were generated using two-tailed Mann–Whitney tests comparing within individual populations and for d using the Kruskal–Wallis test with Dunns correction.