DEV02162 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Neural-specific Nf^hGFAPKO mutant mice are small and have decreased survival. (A) Weight plots of newborn (P0.5), P8 and adult control (CTR) and mutant (CKO) mice. The number of mice was analyzed for each group: P0.5, CTR, n=25, CKO, n=6, P=0.34; P8, CTR, n=17, CKO, n=7, ***P<0.0001; adult male, CTR, n=22, CKO, n=11, ***P<0.0001; adult female, CTR, n=21, CKO, n=14, ***P<0.0001. (B) Survival curves of Nf1^hGFAPKO mutant and control mice. (C) A representative of a control and a Nf1^hGFAPKO mutant adult brain. Arrows indicate the cerebellum (marked by broken lines), which is smaller in the mutant. Scale bar: 1 mm.

• Supplemental Figure 2 -

  Fig. S2. Nf1^GFAPKO mice display extensive astrogliosis in the both gray and white matter of the adult brain. Serial coronal sections were cut at 50 mm and stained with anti-GFAP. Four sections (from rostral to caudal) of control (A-D) and mutant brain (E-H) are shown. High magnification of the control brain (I-L; the boxed area in A-D) and mutant brain (M-P; the boxed area in E-H) are shown. In addition to the gray matter, the mutant white matter such as corpus callosum (M) and anterior commissure (N) has a conspicuous increase in the number of GFAP-positive astrocytes as compared with controls (I,J). Scale bar: 1 mm in A-H; 100 mm in I-P.

• Supplemental Figure 3 -

  Fig. S3. Comparison of two GFAP-cre transgenic strains. Sections of the proximal optic nerve and hippocampus from 2-month-old mice with the genotype of GFAP-cre*+ (A,C) and GFAP-cre*+;Rosa26-lacZ/+ (B,D) were subjected to X-gal staining. The GFAP-cre* transgenic strain has a lacZ transgene under the control of the GFAP promoter (A,C). Cre-mediated recombination in hippocampal neurons is revealed by the Rosa26-lacZ (D), but not by the GFAP-cre*/lacZ (C). The cytoplasmic staining of the Rosa26-lacZ is qualitatively different from the nuclear lacZ staining driven by the GFAP promoter. Sections from the retina with the genotype of GFAP-cre*+;Rosa26-lacZ/+ were stained with anti-NeuN (red) and anti-lacZ (green) (E) or with anti-GFAP (red) and anti-lacZ (green) (F), confirming the astrocytic specificity of the GFAP-cre* transgene in the retina. Scale bar: 100 mm.