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Files in this Data Supplement:
Fig. S1. Expression patterns of Dll1 and active Notch1 in the neocortical ventricular zone. (A-G) E13.5 ICR mouse embryos were excised 30 minutes after BrdU injection. The brain sections were immunostained with anti-BrdU together with anti-active Notch1 (actN1) (A-C) or anti-Dll1 (D-F), or with anti-actN1, anti-Dll1 and TO-PRO-3 (for nuclear staining) (G). Note that most of the cells labeled with BrdU after a 30-minute pulse labeling (cells at around S phase) in the VZ were positive for actN1 and negative for Dll1 immunostaining (A-F), that some BrdU-positive cells in the VZ exhibited weak immunostaining for actN1 and expressed Dll1, and that, although Dll1 expression was enriched in dividing cells located at the ventricular surface, which were mostly devoid of actN1, some cells located at the ventricular surface were positive for actN1 and negative for Dll1 immunostaining (G, and data not shown). (H) The percentage of actN1-positive cells among Dll1-negative or Dll1-positive cells in each region of the VZ of the neocortex (the apical half, the basal half, and the total VZ) was determined by immunohistochemistry. Data are the mean + s.e.m. of values from four sections of each brain, and similar results were obtained from three independent brains. *P<0.01. NCX, neocortex; VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone. Scale bars: 50 µm.
Fig. S2. Effects of Dll1 overexpression on clone size in a clonal assay. (A) Average clone size was determined from the clonal analysis indicated in Fig. 3B. (B) Frequency of clones according to the clone size (number of cells per clone, x-axis) was determined in the clonal analysis in Fig. 3B. (C) The percentage of TuJ1-positive cells at each clone size was determined from the clonal analysis in Fig. 3B. Note that Dll1 expression in a small subset of cells had little effect on the average clone size (A) and the distribution of the clone size (B) in this assay.
Fig. S3. DICD does not promote neuronal differentiation in vivo. DICD was overexpressed by retroviral infection in vivo in the same experiments as indicated in Fig. 5. The distribution of GFP-positive cells in the neocortex was determined. Data are the mean ± s.e.m. of values from eight sections of each brain compared between littermates. Similar results were obtained from five independent brains. *P<0.01, **P<0.0001. VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone; CP, cortical plate.
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