Sfrp5 regulates hepatoblast formation in zebrafish.

<p>Embryos were injected with 0.5 pmol morpholino against <i>sfrp5</i> (MO; B, G, L, Q), 0.5 pmol of the control morpholino (mmMO; A, F, K, P), 100 pg <i>mCherry</i> mRNA (C, H, M, R), or 100 pg <i>sfrp5</i> mRNA (D, E, I, J, N, O, S, T). <b>A</b>–<b>O</b>) Whole-mount <i>in situ</i> hybridization with a probe against <i>hhex</i> staining embryos at 25 hpf (A–E), 36 hpf (F–J), and 48 hpf (K–O). Dorsal view, anterior to the left. Arrowheads point to the hepatoblast. <b>P</b>–<b>T</b>) Confocal microscopy of injected gutGFP embryos. Ventral view with anterior to top. Liver is outlined in white, pancreas in yellow. The scale bar is equal to 25 µm. <b>U</b>) RT-PCR of morpholino and control injected embryos with primer pairs detecting <i>sfrp5</i> or β-actin. <b>V</b>) Bar chart representing distribution of normal and abnormal embryos processed by <i>in situ</i> hybridization, with representative samples shown in A–O. <b>W</b>) Boxplot showing liver size distribution in injected embryos. **: <i>p</i><0.001. <b>X</b>) Boxplot showing distribution of the number of GFP⁺ liver cells. *: <i>p</i><0.05, **: <i>p</i><0.01. <b>Y</b>) Boxplot showing liver cell size distribution. Numbers below each column or boxplot show how many embryos were analyzed.