DEV033498 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Conditional ablation of Brca1 results in a reduced number of deep-layer, but not upper-most layer, neurons in the P8.5 neocortex. (A) Double immunofluorescence (6-µm optical sections) for the upper-layer marker Brn1 (green) and the deep-layer marker FoxP2 (red) on Vibratome sections of P8.5 neocortex of Brca1 control (top) and conditional knockout (cKO, bottom) mice. Sections were consecutive to the Brn2-stained and Tbr1-stained sections shown in Fig. 2B. Cortical layers are indicated by I-VI. Scale bar: 100 µm. (B) Illustration showing the distribution of Brn1⁺ FoxP2⁻ cells (green dots), Brn1⁻ FoxP2⁺ cells (red dots) and Brn1⁺ FoxP2⁺ cells (yellow dots), as revealed by the double-immunofluorescence images shown in A. (C) Quantification of Brn1⁺ FoxP2⁻ cells (green), Brn1⁻ FoxP2⁺ cells (red) and Brn1⁺ FoxP2⁺ cells (yellow), in a 300-µm-wide segment of the cortical wall as shown in A. Data are the mean of three brains (one field per brain); error bars indicate s.e.m.; *P<0.05.

• Supplemental Figure S2 -

  Fig. S2. Conditional Brca1 ablation in the dorsal telencephalon does not alter the abundance or organization of calretinin-positive neurons in the neocortex. Immunofluorescence (6-µm optical sections) for calretinin on P4.5 Vibratome sections of the neocortex of control (Brca1^loxP/loxP Emx1-Cre^−/−, left) and Brca1 cKO (Brca1^loxP/loxP Emx1-Cre^+/−, right) mice. Sections were consecutive to the Nissl-stained sections shown in Fig. 1E′,F′. The marginal zone is at the top of the images. Scale bar: 100 µm.

• Supplemental Figure S3 -

  Fig. S3. Conditional ablation of Brca1 results in the apoptosis of Pax6-positive cells. Double immunofluorescence (1-µm optical section) for Pax6 (green) and nestin (green), together with TUNEL (red) and DAPI (white) staining, of a coronal cryosection of E11.5 dorsal telencephalon of a Brca1 cKO mouse. The top of the image is the VZ-preplate boundary. A TUNEL-positive Pax6-negative cell is indicated by the red arrowhead, and a TUNEL-positive Pax6-positive cell is indicated by the yellow arrowhead. Scale bar: 20 µm.