Fig. 8
Identification of Foxg1 upstream regulators in projection subtype selection. a Schematic diagram of temporal precursor cell labeling using Neurog2CreER/+ and R26LSL-tdTomato mediated genetic recombination upon 4OHT administration. b Schematic diagram of sample collection. Neurog2CreER/+; R26LSL-tdTomato mice were administered with 4OHT at E13.5 or E15.25, and brains were collected at the indicated time points. c Heatmap representing hierarchical clustering using the complete linkage with Euclidean distance. Datasets were obtained from two independent analyses normalized to Z-score and shown in average. Genes with significant P value less than 0.01 by differential expression on edgeR and with enriched expression in E13.5 or E15.5 4OHT administration were selected. d Ingenuity Pathway Analysis comparing cells from E15.25 and E13.5 4OHT treatment at 24 and 48 h. Candidate upstream regulators related to intercellular signaling pathway with P value less than 10−3 are shown. Positive and negative values of Z-score represent activated or repressed state, respectively. Threshold of Z-score = 1 and −1 is marked in gray. e Schematic diagram indicating the 1.3 kb Foxg1 promoter positioned at chr12:50,483,149–50,585,435 on the UCSC genome (NCBI37/mm9). f Quantitative analysis of transcription factors with multiple predicted binding sites on the same strand of the Foxg1 gene (http://tfbind.hgc.jp). g–h′ Expression of Egr1 and Egr2 mRNA by in situ hybridization in E16.5 cortices. i Schematic diagram of in utero electroporation. Brains were introduced with pCAG:GFP and pCAG:empty (Control) or pCAG:GFP and pCAG:Egr1 (Egr1 GOF), or pCAG:GFP and pCAG:Egr2 (Egr2 GOF). Dashed lines indicate the ventricular surface. j–l′ Double immunohistochemistry of GFP (green) and Foxg1 (red) in E16.5 cortices. m Quantitative analysis of the percentage (±SEM) of GFP cells that express Foxg1. n–v Immunostaining of P7 cortices with GFP (green), Satb2 (red) (n, q, t), Rorβ (red) (o, r, u), and vGlut2 (red) (p, s, v) antibodies. w, x Quantitative analysis of the percentage (±SEM) of GFP cells that express Satb2 (w) or Rorβ (x). VZ ventricular zone, IZ intermediate zone. * indicates P value <0.05 and ** indicates P value <0.01 by Student’s t-test. Source data are provided as a Source Data file
