DEV01204 Supplemental Figures

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Comparison of Sox2 mRNA (top, blue) and protein (lower panels; brown, DAB reaction) expression between wild type (wt) and Sox2^b-geo/DENH compound heterozygous mutant (MUT). (Top two panels) E12.5 lateral ventricle (LV). (Third panel) adult lateral ventricle. Asterisks indicate the origin of the rostral migratory stream in the wild type; note the decreased staining of the corresponding region in the mutant. (Bottom panel) Adult thalamus.

• Supplemental Figure 2 -

  Fig. S2. Sox2 mRNA expression in neural precursors grown in vitro from normal and mutant adult brains, analyzed by RT-PCR. Neurospheres were grown for three passages from dissected periventricular zone cells from the brains of adult mice with the indicated genotypes. Total RNA from 100,000 cells was pre-treated with DNAseI, reverse transcribed and amplified with primers for Sox2 and for HPRT, according to Zappone et al. (Zappone et al., 2000). Equal amounts of cDNA from control wild type (+/+), compound heterozygotes (b-geo/DENH) and b-geo/+ heterozygotes were amplified in parallel. In addition, dilutions of the wild-type cDNA (0.2, 0.3, 0.5 and 0.75, as indicated) were also amplified to provide a quantitative reference. Controls ‘retrotranscribed’ in the absence of reverse transcriptase (RT–) were also carried out to rule out genomic amplification. A representative experiment is shown. (A) Ethidium bromide staining at 30 cycles of amplification. (B) Southern blot with Sox2 probe of samples amplified for 20 cycles. The radioactivity was quantitated by analysis with Typhoon 8600 apparatus using ImageQuant software. Band intensity of the wild-type sample was taken to represent 100 units; corresponding values for samples from mutant mice are shown.

• Supplemental Figure 3 -

  Fig. S3. Double immunofluorescence analysis of coronal sections of lateral ventricle from adult brain, with antibodies against SOX2 and PSA-NCAM (confocal microscopy). The section in B is at the anterior extremity of the lateral ventricle, those in A and C are at an intermediate position. For the morphology in C, see Doetsch et al. (Doetsch et al., 1997).