Extended Data Fig. 4: Characterization of Sox2-GFP⁺ and Sox1-GFP⁺ pNSCs.

a, NSC-like cluster forming efficiencies from sorted adult and postnatal lung Sox2-GFP⁺ cells. The data represent means ± s.d. (n = 3 biological replicates). b, Expression levels of NSC marker genes in different samples quantified by RT-qPCR. All data are calibrated to brain NSCs, whose expression is considered to be 1 for all genes. The data represent means ± s.d. (n = 3 biological replicates). c, Lung pNSCs from postnatal Sox2-GFP mouse could differentiate into neurons (Tuj1⁺), astrocytes (GFAP⁺) and oligodendrocytes (MBP⁺) in vitro. Scale bar, 50 μm. d, Dot plot showing the top 4 markers for each cluster in postnatal Sox2-GFP⁺ lung cells. Dot size represents percentage of cells where the gene is detected, colour indicates average expression level of the gene in each cluster. n = 1 biological replicate. e, Immunohistological analysis of lung tissue from adult WT mouse using antibody against Sox1, Sox2. Data represent 3 biological replicates. Scale bar, 50 μm. f, Immunohistological analysis of heart tissue from postnatal WT mouse using antibody against Sox1. Data represent 3 biological replicates. Scale bar, 50 μm. g, FACS strategy for postnatal DRG GFP⁺ and GFP^- cells from Sox1-GFP mouse, and morphology of a primary DRG pNSC cluster and GFP^- cells after culture, as assessed by bright-field (BF) microscopy and GFP signal. h, Primary and passage 1 brain NSCs from postnatal Sox1-GFP mouse, as assessed by bright-field (BF) microscopy and GFP signal. Data represent 3 biological replicates. i, Immunofluorescence microscopy images of lung pNSCs, tail pNSCs and brain NSCs from postnatal Sox1-GFP mouse using antibodies against Sox1, Sox2, Olig2 and Nestin. j, Lung and DRG pNSCs, and tail pNSCs from postnatal Sox1-GFP mice could be stably maintained for more than 50 passages in monolayer. Data represent 3 biological replicates. k, Expression levels of NSC marker genes in different samples quantified by RT-qPCR. All data are calibrated to brain NSCs, whose expression is considered to be 1 for all genes. The data represent means ± s.d. (n = 3 biological replicates). l, Lung, tail and DRG pNSCs and brain NSCs from postnatal Sox1-GFP mouse could differentiate into neurons (Tuj1⁺), astrocytes (GFAP⁺) and oligodendrocytes (MBP⁺) in vitro. m, The in vitro differentiation efficiencies of pNSCs and brain NSCs from Sox1-GFP mouse into neurons, astrocytes, and oligodendrocytes were quantified and compared via immunostaining with Tuj1, GFAP, and MBP, respectively. Brain control NSCs were used as a positive control for determining the relative differentiation efficiency. The data represent means ± s.d. (n = 3 biological replicates). n, Sox1-GFP⁺ cells without expansion with growth factors EGF and FGF-2 could differentiate into neurons, astrocytes and oligodendrocytes in vitro. White arrows indicate the MBP⁺ oligodendrocytes. Data represent 3 biological replicates. Scale bar, 50 μm.