XB-IMG-157849
Xenbase Image ID: 157849
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Fig. 5. FoxD1 patterns mesoderm downstream to BMP antagonism. (A) Embryos were injected marginally at the one-cell stage with mRNA encoding foxd1 (25 pg) protein. VMZ explants were removed from control and injected embryos at early gastrula stage 10.25 and grown to neurula stage 17. Total RNA was isolated from five control embryos (lane 2) and eighteen VMZs from each group (lanes 3-4). Various mesoderm markers along the DV axis were examined by sqRT-PCR: dorsal/notochord (chd, xnot), dorsal/lateral muscle (myod, MA) and ventral (vent1). (B) Embryos were co-injected marginally at the one-cell stage with the FoxD1-MO (25 ng) or mRNAs encoding the foxd1 (25 pg), noggin (20 pg), or dkk1 (35 pg) proteins. VMZ or DLMZ explants were removed from control and injected embryos at early gastrula stage 10.25 and grown to neurula stage 17. Explant elongations were scored as previously described, from three independent experiments for all explant groups: control VMZ (n=42), VMZ-FoxD1 (n=51), VMZ-Noggin (n=51), VMZ-Noggin + FoxD1-MO (n=52), VMZ-Dkk1 (n=51), VMZ-FoxD1 + Dkk1 (n=27), control DLMZ (n=54), DLMZ + FoxD1-MO (n=47). (C) Embryos were injected marginally at the one-cell stage with the FoxD1-MO (25 ng). VMZ explants were removed from control and FoxD1 morphant embryos at gastrula stage 10.25 and VMZs were cultured in soluble noggin to neurula stage 17. Total RNA was isolated from six control (lane 2) and FoxD1 morphant embryos (lane 3), and from eighteen VMZs from each group (lanes 4-7). Expression of dorsal, dorsal-lateral, and ventral markers was examined by sqRT-PCR: chd, xnot, foxd1, myod, MA and vent1. odc serves as a control for quantitating RNA levels. (D) Embryos were injected marginally at the one-cell stage with the FoxD1-MO (25 pg) or mRNA encoding noggin protein. DLMZ explants were removed from control and injected embryos at gastrula stage 10.25, and DLMZs were cultured in soluble noggin to neurula stage 17. Total RNA was isolated from six control embryos (lane 2), and from eighteen DLMZs from each group (lanes 3-5). Expression of the anterior (nkx2.5), dorsal (xnot, chd), and dorsal lateral (MA) markers was examined by sqRT-PCR. Image published in: Polevoy H et al. (2017) Copyright © 2017. Reproduced with permission of the Publisher, University of the Basque Country Press. Larger Image Printer Friendly View |