Extended Data Fig. 2: Characterization of pNSCs derived without low-pH treatment.
a, 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). b, Representative morphology of Nes-GFP+ cells after culture, as assessed by bright-field microscopy. c, Lung pNSCs, tail pNSCs, brain control NSCs and mouse ESCs were injected into SCID mice. Only ESCs, but not lung pNSCs, tail pNSCs and brain NSCs, formed teratomas 4 weeks after injection. d, Nes-GFP+ lung pNSCs, tail pNSCs and brain NSCs were labelled with CAG-H2B-Tomato. Data represent 3 biological replicates. e, Immunohistological analysis of the transplanted cells 6 weeks after transplantation using antibodies against Tomato, Ki67 and Nestin. Data represent 3 biological replicates. Scale bar, 50 μm. f, Genotyping of Cre recombinase by polymerase chain reaction (PCR) in tail pNSCs from WT mouse. g, Postnatal WT mouse tail pNSCs were labelled with CAG-H2B-GFP. Data represent 3 biological replicates. h, The genetic approach to trace cell fusion between transplanted cells and endogenous cells. i, The transplanted tail pNSCs labelled with CAG-H2B-GFP were RFP- and differentiated into mature neurons (NeuN+/GFP+), astrocytes (GFAP+/GFP+), and oligodendrocytes (MBP+/GFP+). White arrows indicate the differentiated neurons, astrocytes and oligodendrocytes. Data represent 3 biological replicates. Scale bar, 50 μm.
