The Journal of Neuroscience, October 19, 2005, ():
Role of Fabp7, a Downstream Gene of Pax6, in the Maintenance of Neuroepithelial Cells during Early Embryonic Development of the Rat Cortex
J. Neurosci. Arai et al.
25: 9752
Supplemental data
Files in this Data Supplement:
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Supplemental material.
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Supplemental Fig. 1. Down-regulation of Fabp7 gene in rSey2/rSey2 rat brains. (A) Scatter plot analysis of genes differentially expressed in the FB and HB samples at E12.5. The vertical and horizontal lines indicate the expression levels in rSey2/rSey2 and the wild type (WT), respectively. Genes with more than 2.0 fold changes (squares above and below lines +/- 2) are considered as up- and down-regulated, respectively, in rSey2/rSey2 brain samples. Fabp7 is extremely down-regulated both in the FB and HB fractions. (B) Down-regulation of Fabp7 is confirmed by quantitative real-time PCR using LightCycler. Data are the fold changes in microarray (gray bar) and results of quantitative PCR (black bar).
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Supplemental Fig. 2. Secondary structure and down-regulation of Fabp7 mRNA. (A) Prospective secondary structure of Fabp7 mRNA created by MFOLD program. Red brackets show the recognition sites of five types of small hairpin RNAs (shRNAs) (i-148, i-149, i-246, i-247 and i-274). (B) Sequences of five shRNAs. The name of each construct was made by nucleotide positions of Fabp7 mRNA. (C) Reduction of Fabp7 mRNA expression using i-149 detected by whole mount in situ hybridization. Rat embryos were electroporated with pSUPER/i-149 at E11.5 and cultured for 24 hours by whole embryo culture. Red brackets show the electroporated sites and reduction of Fabp7 mRNA expression compare to the control side. Left and right panels show the rostral and caudal sides of telencephalon.
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Supplemental Fig. 3. A schematic model of Pax6-induced regulation of NEp cell proliferation and differentiation during early development of the cortex. At the early stage, Pax6 predominantly transcribes Fabp7 in proliferating NEp cells and maintains their epithelial morphology (A). As development proceeds, Pax6 switches on the expression of Ngn2 in some neuronal-committed NEp cells in which Fabp7 is still transcribed (B). Apical progenitor cells appear in which Ngn2 is dominantly transcribed by Pax6 (C). At later stages, basal progenitors emerge in which Ngn2 transcription is independent of Pax6 (D).
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Supplemental table 1.
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Supplemental table 2a.
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Supplemental table 2b.
