pSMAD nuclear translocation defects in ipo8?/? embryos highlight impaired TGF-?/BMP signaling (A) Maximal projections of confocal Z stacks of WT and ipo8?/? embryos labeled for pSmad1/5/9 (green) and DAPI (blue) showing well-defined ventral to dorsal gradient of pSmad5 in WT gastrulating zebrafish embryos (left panel) but not in the ipo8?/? embryos (right panel). Scale bar represents 100 ?m. (B) Confocal images of the ventral sides of WT and ipo8?/? embryos labeled for pSmad1/5/9 (green), phalloidin (red), and DAPI (blue). Arrowheads highlight the membrane localization of the pSmad staining in the ipo8?/? mutants. Scale bar represents 20 ?m. Dot plots representative of three experiments show quantifications of the ratio of pSmad1/5/9 cytoplasmic signal over nuclear signal; 5 WT and 6 ipo8?/? embryos were analyzed. Median and IQR are shown. p values were calculated by Mann Whitney test (??p < 0.01). (C) Heatmaps of differentially expressed genes between WT and ipo8?/? embryos at 13 and 24 hpf. Three to four biological replicates are shown per group. The top enriched gene ontology term for biological process is highlighted for each cluster.
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