kremen1 Is Present in the Endoderm at 24 hpf, and Its Roles in Early Endoderm and Liver Development. related to Figure 4 (A) kremen1 (krm1) is ubiquitously expressed at 24 hpf. (B and C) Endodermal cells are collected by sorting out GFP positive cells from Tg(sox17:GFP) transgenic embryos. (D) Presence of kremen1 in the endoderm is confirmed by RT-PCR using sorted endodermal cells. Transcriptions of foxa3 and otx2 are applied as positive and negative controls, respectively. (E and F) In contrast to the wild-type (E), injection of kremen1 mRNA results in slightly increased number of endodermal cells and mildly expanded endoderm at the 14-somite stage (F) as observed under the Tg(sox17:GFP) background. (G and H) Injection of kremen1MO leads to reduced number of endodermal cells (G), which is rescued by the co-injection of kremen1 mRNA (H). (I and J) Expressions of Kremen1-GFP at 26 hpf caused by heat shock of Tg(hsp70l: kremen1-GFP) transgenic embryos at 18 hpf. (K and L) In contrast to the Kremen1-GFP negative embryos (K), liver development is aberrant in the Kremen1-GFP positive embryos at 102 hpf (L).
|