Fig. 3

Generation and characterization of SMASh tagged FOXG1 hESCs. a Strategy for generation of FOXG1^s/s hPSC lines using CRISPR/Cas9. b Sanger sequencing of DNA fragment of a representative FOXG1^s/s hESC colony showing the integration of HA-SMASh. c, d Immunofluorescence images (c) and quantification (d) (n = 5 experimental replicates) of cortical neuronal progenitors derived from FOXG1^s/s hESCs for FOXG1 (red) and HA (green). e–g Western blotting for FOXG1 protein of WT hESC-derived NPCs (e), FOXG1^s/s hESC-derived NPCs (f) and FOXG1-GGS hESC-derived NPCs (g) under the treatment of ASV (0, 50, 100 , and 100 nM). h, i Western blot (h) and intensity analysis (i) (n = 3 experimental replicates) for FOXG1 protein of FOXG1^s/s hESC-derived NPCs under treatment with different concentration of ASV (0, 10, 25, 50, 75, 100, 300, and 1000 nM). j Immunofluorescence images of FOXG1-expressing telencephalic progenitors for HA (green), DAPI (blue) upon treatment with different concentration of ASV (0, 50, 100, and 1000 nM), showing dosage regulation of FOXG1 protein. All error bars represent mean ± s.e.m. All scale bars 100 μm. Source data are provided as a Source Data file