DEV02290 Supplementary Material

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• Supplemental Figure 1 -

  Fig. S1. Characterization and birth date analysis of GFP-labeled cells. (A¢¢) Immnostaining with anti-glutamate antibody in GFP-labeled embryos. Migrating GFP-positive cells expressed glutamete (arrows). (B-B¢¢) Immunostaining with anti-BrdU antibody in GFP-labeled embryos. Embryos were cultured in the medium containing 20 mg/mg of BrdU for 30 minutes, immediately after GFP-plasmid electroporation. After washing out BrdU, the embryos were transferred into fresh medium and further cultured for 48 hours. BrdU was incorporated into the GFP-positive cells that aligned at the PSB (arrows). Scale bars: 200 mm in A; 20 mm in B.

• Supplemental Figure 2 -

  Fig. S2. Distribution of GFP-positive cells in transplantation experiments. (A-D) Coronal sections of wild type (donor: Pax6 mutant) (A,B) and Pax6 mutant (donor: wild type) (C,D) telencephalon. To verify the PSB, in situ hybridization with Pax6 probe was performed on serial sections (B,D). In wild-type telencephalon, Pax6 mutant-derived cells stopped at the PSB, whereas in the mutant telencephalon (A,B), wild-type-derived cells crossed the PSB and invaded the ventral part of the telencephalon (C,D). Scale bar: 200 mm.

• Supplemental Figure 3 -

  Fig. S3. Expression of ephrin-A5 in the ventral part of the telencephalon. In situ hybridization with ephrin A5 (A), Pax6 (B), Dlx1 (C) probe was performed in serial sections of E12.5 wild type rat telencephaolon. At this stage ephirn A5 was specifically expressed at the Dlx1-positive ventral part of the telencephalon. Open arrows indicate the PSB. Scale bar: 100 mm.

• Supplemental Figure 4 -

  Fig. S4. Induction of ephrin A5 expression in Pax6 mutant by misexpression of Pax6. Pax6 expression vector is electroporated into the Pax6 mutant telencephalon. Expression of Pax6 in the ventral part of the telencephalon is detected with anti-Pax6 antibody (A). ephrin A5 expression is induced in the ventral part of the telencephalon, where the Pax6 expression vector is electroporated (B). Scale bar: 100 mm.

• Supplemental Figure 5 -

  Fig. S5. Normal development of the PSB radial glial fibers in the ephrin A5^–/– mouse. (A,B) Expression of reticulon 1 (Rtn1), a marker of the PSB radial glial fibers, in ephrin-A5^+/– (A) and ephrin-A5^–/– (B) mice. The reticulon 1-positive PSB radial glia fibers normally develop in ephrin A5^+/– as well as that in ephrin A5^+/– (arrows in A,B). Scale bar: 100 mm.