Extended Data Fig. 3: Ex vivo and in vitro evaluation of Lrig1 progeny fitness after wound experience.
From: Tissue memory relies on stem cell priming in distal undamaged areas
a, Venn diagram of differentially expressed genes across the time course between Lrig1 and Lgr5 GL cells. b, Quantification of Ki67, as ratio to 0w. Dashed line represents 0w mean. n = 3 (0w, 1w pw1, 1w pw2), n = 4 (8w pw1). c-j, Ex vivo and in vitro experimental settings at 8w pw1, as in Fig. 2c: Lrig1 GL skin biopsies from wound-educated (Distal- 2 to 7 mm from wound bed) and control areas (Ctrl- > 2 cm from wound bed). c, Ex vivo assay assesses migratory ability for epidermal Krt14+ (green) cells. d, Percentage of Ki67+ nuclei. Epidermal cells exiting skin explants do not proliferate. Data are mean ± SEM. n = 3. e, Exit area (mm2) of Lrig1 GL cells from the explant, related to Fig. 2d. Data are mean ± SEM. n = 9. f, In cultured cells Krt6 positivity is inversely proportional to cell migration82. Percentage of Krt6+ in Lrig1 GL cells (left) and representative pictures (right). n = 4. g, Cis-Golgi (Gm130) distribution in Distal vs Ctrl cells. Percentage of polarised cells (marked by asterisk) in GL tdTomato+ and tdTomato- cells (left) and representative pictures (right). n = 8 (Ctrl), n = 10 (Distal). h, Speed of migration (μm/s) of Lrig1 GL cells from in vitro time lapse migration assay, relative to Fig. 2e, f. n = 4. i, j, Percentage of adhesion (i) and survival (j) of Lrig1 GL cells isolated from Distal or Ctrl HFs. n = 4 mice. k, Bulk RNA-seq of Lrig1 GL cells from Distal or Ctrl area after 7 days of culture. Heatmap of deregulated genes is shown. n = 8 (Ctrl), n = 6 (Distal). l, Pie charts illustrating the proportions of the memory genes subsets from RNA-seq of Lrig1 and Lgr5 GL cells in vivo, related to Fig. 2a. m, LogFC heatmap shows memory genes at each time point with respect to 0w. Constitutive (green) and inducible (yellow) memory genes, related to Fig. 2a. n = 3 mice. Data are median with 25th and 75th percentiles. P-value (P) from a two-tailed t-test. Data are mean ± SD, if not differently indicated. Scale bars: 50 μm (b, e); 20 μm (g).
