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Files in this Data Supplement:
Fig. S1. Expression pattern of Cath5, Cash1 and neurogenin 2 (Ngn2) in the central region of E7.5 retina electroporated with control and HA-Hairy1-ΔW-VP16-expressing RCAS provirus. Although the dominant-negative form of chick Hairy1 did not increase neuronal differentiation in the central region of the retina that normally does not express Hairy1 (see Fig. 5D), the ectopic expression of Hairy1 in this region did suppress neuronal differentiation (see Fig. 4A). Therefore, it is theoretically possible that the same target neurogenic genes suppressed by the exogenous Hairy1 could be abnormally upregulated by the dominant-negative Hairy1 with its VP16 activator domain, activation that should lead to an increase in neuronal differentiation even in the central region of the retina. To confirm whether this was the case, we examined the expression pattern of proneural genes (Cath5, Cash1 and Ngn2) that are negatively regulated by Hairy1. Note that the dominant-negative molecule does not change the expression of these proneural genes, suggesting that they are already maximally upregulated in the cells in which the target sites for Hairy1 are available.
Fig. S2. Hierarchical clustering of genes with higher expression in Wnt2b-expressing retina than in control or Delta1-expressing retinas. Data from two independent experiments are shown. Red indicates an expression level above mean expression of a gene across all samples, white represents mean expression, and blue represents expression lower than the mean. The ten transcription factors that we studied and the CMZ marker genes are labeled in yellow and green, respectively. For further information, see Table S1 in the supplementary material.
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