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gata4xenopus   

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Experiment details for gata4

Timing is everything: Reiterative Wnt, BMP and RA signaling regulate developmental competence during endoderm organogenesis.

Timing is everything: Reiterative Wnt, BMP and RA signaling regulate developmental competence during endoderm organogenesis.

Gene Clone Species Stages Anatomy
gata4.S laevis NF stage 37 and 38 heart , endoderm , midgut , foregut

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  Fig. 4. Wnt/BMP-mediated A-P patterning impacts developmental competence. (A,B) Experimental diagram. Xenopus embryos at the 16-cell stage with clear pigment differences (animal pole, dorsal-anterior, and ventral-posterior views of such embryos are shown) were injected with 25 pg of Noggin or 100 pg of Dkk1 RNAs (to inhibit BMP and Wnt/βcatenin signaling, respectively) into each ventral-vegetal V2.1 blastomere along with 25 pg eGFP RNA as a lineage tracer, which targets the ventx1/2-expressing posterior mesendoderm. Hhex+ AE tissue was targeted by injection of each dorsal-vegetal D1.1 blastomere with 25 pg eGFP RNA. At gastrula stage NF10.25, GFP fluorescence was monitored and used to dissect AE, PE, and BMP- or Wnt-inhibited PE explants, which were then treated as indicated in panel B (50 nM RA from NF14-25 followed by 3.5 μM Bio + 50 ng/mL BMP4 from NF25-38). (C)In-situ hybridization analysis of hhex and ventx1+2 (both probes mixed together) in bisected gastrula NF10.25 embryos confirms effective inhibition of the posteriorizing Wnt and BMP pathways by dkk1 and noggin RNA injection. Numbers in the lower left corner indicate numbers of embryos assayed with the gene expression pattern shown. (D-M) Relative gene expression analysis (RT-qPCR) of different anterior-posterior endoderm lineages assayed in explants as prepared/treated in panels A,B. Graphs display the average 2−δδCt value +/- SEM of 3 biological replicates. (N-W)In-situ hybridization of control embryos showing the endogenous spatial domains of expression along the anterior-posterior axis.