Extended Data Fig. 2: Distal memory is specific to the Lrig1 GL lineage.
From: Tissue memory relies on stem cell priming in distal undamaged areas
a-e, IL-17-expressing γδ T cells, responsible for epidermal mTOR and HIF1α induction during wound healing48, are recruited in wound, but not in distal memory areas, at the two acute healing phases 1w pw1 and 1w pw2, while absent at 8w pw1. a, Gating strategy for T cells (CD3+) and γδ T cells (CD3+ γδTCR+) among CD45+ cells and IL-17. b, c, Flow cytometry quantification (%) of T cells (b) and γδ T cells (c) following the scheme in Extended Data Fig. 1l. n = 2 (1w pw2), n = 3 (1w pw1, 8w pw1), n = 5 (0w) mice. d, Staining of CD3 (green) in skin sections. e, IL-17 intensity in T cells quantified by flow cytometry. n = 2 (1w pw2), n = 3 (1w pw1, 8w pw1), n = 5 (0w) mice. f, Epidermal whole-mount highlighting the removal of Lrig1 progeny-derived IFESCs (red), before and after the second overlapped wound. g, Quantification of first (1w pw1) and second (1w pw2) wound closure as distance from wound centre, relative to Fig. 1c. n = 8 (Lgr5), n = 10 (Gata6), n = 14 (Lrig1). Data are mean ± SEM. h, 3D surface plot of Lrig1 GL cells at 0w and 1w pw1, 8w pw1 and 1w pw2, 5 mm from wound bed. Dashed line represents the IFE-HF boundary. Yellow arrows indicate wound bed (Wb). i, Representative confocal images of Lrig1 GL cells (red) exiting toward IFE wound bed at 1w pw1, 8w pw1 and 1w pw2. j, Epidermal whole-mounts (red channel) of GL tdTomato+ cells showing their contribution on the wound bed. Dashed circles highlight the original wound perimeter. k, Number of engaged HFs in Gata6 and Lgr5 GL cells. n = 10. l, m, HFs’ engagement is shown for 1w pw1 and 1w pw2, when the second wound is performed overlapped (1w pw2_Over) or distally (1w pw2_Distal) to the first one. n = 3 (1w pw2_Distal), n = 6 (1w pw1, 1w pw2_Over) (l). Confocal images of engaged HFs showing Lrig1 GL cells (red). Dashed white lines highlight representative engaged HF triplets (m). P-value (P) from a two-tailed t-test. Data are mean ± SD, if not differently indicated. Scale bars: 100 μm (d, h, i); 1 mm (f, j, m).
