ZFIN ID: ZDB-FIG-200817-14
He et al., 2020 - Nanog safeguards early embryogenesis against global activation of maternal β-catenin activity by interfering with TCF factors. PLoS Biology   18:e3000561 Full text @ PLoS Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: 1-cell to Adult
PHENOTYPE:
Fish:
Observed In:
Stage Range: 1-cell to Adult

Fig 5 Transcriptional level of Wnt/β-catenin components and nuclear accumulation of β-catenin are not increased in MZ<italic>nanog</italic>.

(A) WISH and (B, C) RT-qPCR showed the maternal transcription of wnt8a1, wnt8a2, and ctnnb2 were not affected in MZnanog eggs when compared with WT eggs. Scale bar, 100 μm. Error bars, mean ± SD, *P < 0.05; NS means no significant difference. (D) Western blot analysis of total β-catenin and nuclear β-catenin (active β-catenin) in WT, Mznanog, and wnt8a overexpressed embryos. Anti-total β-catenin was used as the β-catenin expression control and anti-β-actin was used as the internal control. A total of 2 pg of wnt8a mRNA was injected at the 1-cell stage in WT and used as a positive control. Embryos were collected at 4 hpf. Experiments were carried out for triplicates. (E) Statistical analysis of active β-catenin/total β-catenin level in panel D. Error bars, mean ± SD, ***P < 0.001. (F) Immunolocalization of β-catenin on whole-mount embryos at the 512-cell stage shows that nuclear β-catenin in both the WT and MZnanog was localized in dorsal margin cells and nondorsal cells, and nuclear β-catenin localization was not stimulated in MZnanog embryos. Signals were observed at animal view. Nuclei were co-stained with DAPI. Scale bar, 50 μm. The P values in this figure were calculated by Student t test. The underlying data in this figure can be found in S1 Data. hpf, hours post fertilization; MZnanog, maternal zygotic mutant of nanog; RT-qPCR, reverse-transcription quantitative PCR; WISH, whole-mount in situ hybridization; WT, wild type.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ctnnb2 WT standard conditions 16-cell whole organism ISH
Adult oocyte ISH
nanogihb97/ihb97 standard conditions 16-cell whole organism ISH
Adult oocyte ISH
nanogihb98/ihb98 standard conditions 16-cell whole organism ISH
Adult oocyte ISH
wnt8a WT standard conditions 1-cell whole organism RTPCR
16-cell whole organism ISH
64-cell whole organism RTPCR
Sphere whole organism RTPCR
Adult oocyte ISH
Adult ovary RTPCR
Adult whole organism RTPCR
nanogihb97/ihb97 standard conditions 1-cell whole organism RTPCR
64-cell whole organism RTPCR
16-cell whole organism ISH
Sphere whole organism RTPCR
Adult oocyte ISH
Adult ovary RTPCR
nanogihb98/ihb98 standard conditions 1-cell whole organism RTPCR
16-cell whole organism ISH
64-cell whole organism RTPCR
Sphere whole organism RTPCR
Adult oocyte ISH
Adult ovary RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab17-ctnnb IHC WT standard conditions 512-cell whole organism nucleus
IHC nanogihb97/ihb97 standard conditions 512-cell whole organism nucleus
IHC nanogihb98/ihb98 standard conditions 512-cell whole organism nucleus
WB WT standard conditions Sphere whole organism nucleus
WB nanogihb97/ihb97 standard conditions Sphere whole organism nucleus
WB nanogihb98/ihb98 standard conditions Sphere whole organism nucleus
Phenotype Details
Fish Conditions Stage Phenotype
nanogihb97/ihb97 standard conditions 1-cell whole organism wnt8a expression decreased amount, abnormal
64-cell whole organism wnt8a expression decreased amount, abnormal
Sphere whole organism wnt8a expression decreased amount, abnormal
Sphere whole organism nucleus Ab17-ctnnb labeling decreased amount, abnormal
Adult ovary wnt8a expression decreased amount, abnormal
nanogihb98/ihb98 standard conditions 1-cell whole organism wnt8a expression decreased amount, abnormal
64-cell whole organism wnt8a expression decreased amount, abnormal
Sphere whole organism wnt8a expression decreased amount, abnormal
Sphere whole organism nucleus Ab17-ctnnb labeling decreased amount, abnormal
Adult ovary wnt8a expression decreased amount, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS Biol.