Fig. 1: Lineage tracing for SOX10 positive cells in postnatal mouse adrenal glands.

A Schematic illustrating that tamoxifen (Tam) induced to the nursing female mouse by gavage in the first 4 days (P0–P4) after she gave birth. The Sox10^CreERT2+/−;R26R^YFP+/− pups induced with Tam by breast-feeding during these 4 days. After 1, 10, or 86 days, the adrenal glands harvested at P5, P14, or P90 for sectioning, staining, and analysis. B Percent stacked bar plot showing the proportion of YFP⁺TH⁺ double-positive cells among total YFP⁺ cells, based on immunofluorescence staining and semi-manual cell counting (P value of P14: 14.17% ± 3.40%, n = 4; P90: 17.56% ± 2.56%, n = 6). Data passed the Shapiro-Wilk normality test (P14: 0.3437; P90: 0.0824), and an unpaired t-test with Welch’s correction revealed no significant difference between P14 and P90 (P value = 0.4544). Numbers above each bar indicate the average total number of YFP⁺ cells per time point. Data are presented as mean ± s.e.m. C Bar plot showing the total number of medullary TH⁺ cells, based on immunofluorescence staining and semi-manual cell counting (P14: 1067.00 ± 75.14, n = 4; P90: 2542.00 ± 128.90, n = 6). Normality was confirmed by Shapiro-Wilk test (P value of P14: 0.3567; P90: 0.6246), and a t-test with Welch’s correction showed a significant increase in TH⁺ cells at P90 compared to P14 (P value < 0.0001). Numbers in each bar indicate sample size (n). Data are presented as mean ± s.e.m. ****, P value < 0.0001. D–F Immunofluorescence staining for the adrenal glands harvested at P5, P14, and P90 after Tam-milk fed. Full arrowheads indicate YFP and TH double-positive (YFP⁺TH⁺) cells. Scale bars in the adrenal gland images and insets are 100 μm and 20 μm, respectively. G H&E and immunofluorescent staining for the same window in (F). Empty arrowheads indicate YFP and SOX10 double-positive (YFP⁺SOX10⁺) cells showing lineage tracing efficiency. Full arrowheads indicate YFP and TH double-positive (YFP⁺TH⁺) cells. Scale bars are 20 μm. H, I Immunofluorescent staining for the adrenal glands harvested at P5, and P14 after Tam-milk fed. Empty arrows indicate EdU and TH double-positive (EdU⁺TH⁺) cells. Scale bars in the adrenal gland images and insets are 100 μm and 20 μm. P, postnatal day; Tam, tamoxifen; d, days; TH, tyrosine hydroxylase; YFP, yellow fluorescent protein; DAPI, 4′,6-diamidino-2-phenylindole; EdU, 5-Ethynyl-2’-deoxyuridine; n, sample number; vs., versus.