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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
DVDY_22211_sm_SuppFig1.tif633KFig. S1. A,B: Cell death assay in a normally developing gastrula embryo (A) and a positive control treated with DNase (B). Very few cell death-positive signals were detected in the normal embryo compared with the positive control. These embryos were fixed with PFA and then examined using the In Situ Cell Death Detection Kit, Fluorescein (Roche) according to manufacturer's protocols. Scale bar = 50 μm.
DVDY_22211_sm_SuppFig2.tif1282KFig. S2. Determination of the numbers of injected epithelial cells. Epithelial cells were stained with 2 μg/ml of fluorescein isothiocyanate (FITC) in the same manner as rhodamine B isothiocyanate-celite (RITC) -staining. Eight hours after injection, the embryos were fixed with paraformaldehyde (PFA) and the numbers of propidium iodide (PI) -stained nuclei were determined using a confocal microscope. A,B: The Nomarski and stacked microscopic images of an epithelial aggregate, respectively. Green; FITC signal, red; PI signal. Scale bar = 50 μm.
DVDY_22211_sm_SuppFig3.tif351KFig. S3. Comparison of 3-day-old bipinnaria larvae derived from embryos injected with different numbers of mesenchyme cells at the blastula stage. The larvae were fixed with PFA and then stained for immunofluorescence analysis using the MC5 Mab, a mesenchyme cell marker, as described previously (Furukawa et al., 2009). A,B: An aboral half stacked image of a larva produced after injection of the moderate number of cells is shown in A, that of the excess number of cells in B. Red, injected mesenchyme cells; green, innate mesenchyme cells. Scale bar = 100
DVDY_22211_sm_SuppTab1.xls36KSupplement Table 1
DVDY_22211_sm_SuppTab2.xls36KSupplement Table 2

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