DEV00615 Supplemental Data

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• Figure S1 - Xhox3 is a direct target of BMP and activin signals. Animal caps injected with mRNA encoding CA-ALK2 (150 pg) or activin (3 pg) were isolated at stage 8 and treated with cycloheximide (CHX). The expression of marker genes was detected by RT-PCR at stage 11. Note that BMP and activin signaling, stimulated by CA-ALK2 and activin, respectively, induced Xhox3 expression even in the presence of CHX, while the expression of chordin, an indirect target of activin, is suppressed by CHX.

• Figure S2 - In vivo distribution of p53 transcriptional-activating activity during gastrulation. A p53 reporter gene, p53 (X3), was injected into different parts of four-cell embryos, which were then cultured to early gastrula stages. Whole embryos were used for the luciferase assay as described in Materials and Methods. A reporter vector containing the Otx minimal promoter without the p53-binding sites was used as a control. The columns indicate the averages of triplicate assays and the error bars indicate the ranges. Two other independent experiments gave similar results.