Fig. 4

Decreased endoderm-derived organ size in copeb morphants is not due to increased cell death. (a) Cell death was examined in live copeb ATG morphants and control embryos at 24, 48, 72 and 96 hpf by acridine orange. There was increased labeling of the head and eye in copeb morphants at 24 hpf but virtually no staining in the control and morphants at any other time. White arrowheads point to the liver, which does not have any acridine positive cells in either morphants or controls. n = 60 for each sample at each time point. ot = otolith. (b) No increase in apoptosis is observed in endoderm-derived organs in 3 and 4 dpf copeb ATG morphants compared to controls. Sections of embryos on 3 and 4 dpf were stained with Cy3-streptavidin to label the liver and gut epithelium, and were processed for TUNEL using fluorescein. TUNEL positive cells (white arrow heads) are observed only in very few cells in 3 dpf sections in the brain of copeb ATG morphants. Blue = DAPI, Red = Cy3 Streptavidin, Green = TUNEL, scale bar = 50 mM.