Fig. 3: Olig2 plays a key role in stem cell fate transition.

a Phase object confluence of adherently grown Ptch1^+/−;Trp53^−/− CRISPR-Cas9 control or Olig2-KO mouse tumour cells measured across 9 days using live cell imaging. The cells were grown in either proliferation media, modified quiescence-inducing media containing BMP4 or modified media control for 3 days and then washed and re-exposed to proliferation media; data shown are representative of three independent experiments; error bars denote mean ± SEM; two-tailed unpaired t-test performed at final time point (212 h); two-tailed p-values; ns p ≥ 0.05, ****p < 0.0001; p = 0.4693 (Olig2-KO + veh vs. control + veh), p = 0.4967 (Olig2-KO + MM vs. control + MM), p < 0.0001 (Olig2-KO + MM + wash vs. control + MM + wash). b, c Expression of endogenous SOX2 and OLIG2 proteins detected by immunofluorescence in the external granule layer (EGL) at P14 in Ptc WT and Ptc mice (b) and quantification of the SOX2⁺/OLIG2⁺ cells as a fraction of the SOX2⁺ cells and OLIG2⁺ cells respectively (c); n = 3 biological replicates; error bars denote mean ± SEM; two-tailed unpaired t-test; *p < 0.05; p = 0.0177. d–k scRNAseq analysis on CGNP-like (SOX2⁺ or DCX⁺) Math1-Cre;SmoM2 mouse tumour cells at P7; unsupervised clustering using Uniform Manifold Approximation and Projection (UMAP) performed on ~5000 single cells (d), dot plot of the top genes most differentially expressed in each cluster (e), graphical demonstration of dimension separation strategy of neoplastic CGNP-like cells (f), construction of a neuronal differentiation trajectory by Monocle2 and the expression of Sox2, Olig2, Dcx, Neurod1, and Stmn2 across the trajectory (g), construction of a cell cycle pseudotime by slingshot and the expression of cell cycle markers across the cell cycle pseudotime (Pcna, Mki67, and Top2a) (h), heatmap of the differentially expressed genes across the neurogenesis trajectory and the cell cycle pseudotime (i), comparison of the neurogenesis and cell cycle trajectories; gene names associated with each heat map are listed in Supplementary Data 2 (j), violin plots of cell cycle pseudotime and Mki67 expression between OLIG2⁺ vs. OLIG2^– cells; centre line, median; box bounds, upper and lower quartiles; whiskers, 1.5x interquartile range; points, outliers; n = 1096/3996 (OLIG2⁺/OLIG2⁻); two-tailed unpaired t-test (k). Source data are provided as a Source Data file.