Fig. 2.

Phenotypic consequences of Sox1 induction. a Sox1^GFP induction can endure prolonged time periods. Sox1^GFP positive cells were FACS sorted from control (no gRNA) and experimental settings (SoxProm), cultured for 7 more days and eventually analyzed by flow cytometry. While Sox1^GFP-positive cells sorted from control populations show no relative enrichment, those from the activated population show a strong increase of the number of cells in the GFP-positive gate after 7 days, indicating some steadiness of the gene induction. b, c Sox1 activates hundreds of genes associated with a stem cell identity. b Heat map showing the 100 genes most significantly induced by Sox1 induction (NPC+). Most of these genes are also higher in the transcriptomes of early neural stem cells (NSCs, 7 days neural induction of ESCs; NR, sorted neural rosette cells). c Relative expression changes of a selected set of genes are shown; data shown as (logarithmic) fold change and standard error of the mean; all data are depicted for three individual biological replicates. d, e Differentiation assays reveal changes in the potency of Sox1-positive NPCs. After gRNA transduction, Sox1^GFP-positive and -negative cells were sorted from both control (no gRNA) and activated (S1-9) populations and differentiated for 7 days; d scale bars (upper row: 20 µm; lower row: 100 µm). e The number of cells positive for the neuronal marker TujI increased significantly in Sox1-positive cells (***p < 0.001 in two-sided Student’s t-test; n = 3 biological replicates, generated independently on different days in different clonal lines)