DEV02557 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. No transpecification is observed in the Mgn^tz/tz-expressing cells. (A-F) In situ hybridization with Gad67 probe in sagittal sections of postnatal tissue on wild-type (Mgn^+/+) and on Mgn^tz/tz brains at P25. The dorsal mesencephalic vesicle and rostral forebrain are magnified in wild type (C,D) and homozygous mutant mice (E,F), respectively. Gad67-expressing cells are absent in the SC and reduced in number in the IC. (G-O) In situ hybridization with Gat1 (marker for GABAergic cells), Vglut1 (marker for glutamatergic cells) and Chat1 (marker for cholinergic cells) probes in sagittal sections showing the mesencephalic vesicle in postnatal brain on wild-type (center column) and on Mgn^tz/tz (right column) mice at P20. (G,J,M) Bright-field images of homozygous mutant mice. Abbreviations: Cb, cerebellum; CPu, caudate putamen (striatum); Cx, cortex; IC, inferior colliculus; OB, olfactory bulb; SC, superior colliculus.

• Supplemental Figure 2 -

  Fig. S2. BrdU injection and TUNEL assays. (A) Images in B-E are taken from the area of the SC of the indicated genotypes corresponding to the area depicted by a square in the bright-field image. (B,C) Distribution of BrdU positive cells in the SC of wild-type (B) and Mgn^tz/tz mutant (C) mice on coronal sections of E12.5 embryos after 10 minutes of BrdU incorporation. (D,E) Analysis of apoptosis by a TUNEL assay on coronal sections of E12.5 embryos. TUNEL-positive cells (green fluorescence signal) were rarely detected in the SC in wild-type (D) and Mgn^tZ/tZ (E) embryos.

• Supplemental Figure 3 -

  Fig. S3. Other neural markers are not disturbed. In situ hybridization with Brn3a (A,B) and Isl1 (C,D) probes at E13.5. In situ hybridization with Th (E-G), Dat (H-J) and Plp1(K-M) probes at P15 in coronal (A,B,E-J), sagittal (C,D) and horizontal (K-M) sections, showing the mesencephalic vesicle on wild-type (Mgn^+/+) and on Mgn^tz/tz tissue. (E,H,K) Bright-field images. No changes concerning these non-GABAergic markers could be observed by comparing wild-type and the mutant mice.