ZFIN ID: ZDB-FIG-161111-2
de Peralta et al., 2016 - Cnbp ameliorates Treacher Collins Syndrome craniofacial anomalies through a pathway that involves redox-responsive genes. Cell Death & Disease   7:e2397 Full text @ Cell Death Dis.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Day 4

Fig. 2

Craniofacial phenotypes generated by tcof1 knockdown were partially rescued by Tp53 depletion. (a) Western blotting showing the Tp53 abundance in control (STD), TRA and TRA+ATGTp53 24 hpf embryos. Molecular weight markers are shown at the right. Tubulin was analyzed as a loading control. The graph shows the densitometric quantification of western blotting band (normalization of Tp53 signal with respect to tubulin). Bars represent mean of relative abundance and S.E.M., n=3, *P<0.05 (t-test) AU: arbitrary units. (b) Expression analysis of Tp53 targets: ccng1 (encoding cyclin G1), pmaip1 (encoding Noxa) and tp53inp1 (encoding tumor protein p53-inducible nuclear protein 1) in control (STD), TRA and TRA+ATGTp53 24 hpf embryos. Bars represent mean of relative abundance and S.E.M., n=3, *P<0.05, (ANOVA test). (c) Acridine Orange staining in vivo performed in 24 hpf embryos. Representative photos of embryos displaying normal (+), intermediate (++) or extensive (+++) cell death. The bar graph shows the distribution of each phenotype among the groups (STD, TRA and TRA+ATGTp53 embryos). ey=eye, n=50 embryos in each of three independent experiments. ***P<0.001 versus STD; ###P<0.001 versus TRA; NS=non-significant P>0.05 (chi-square test). (dh) Quantification of craniofacial parameters (as defined in Figure 1) measured in STD, TRA-MO TRA+ATGTp53 larvae (4 dpf), stained with Alcian Blue. (d) Meckel Area, (e) PQ, (f) Ceratohyal length, (g) CD, (h) Cr. Bars represent means in AU and S.E.M. n=47 for STD, n=79 for TRA, n=30 for TRA+ATGTp53. *P<0.05, **P<0.01, ***P<0.001 versus STD; #P<0.05, ##P<0,01 ###P<0.001 versus TRA; NS=non-significant P>0.05 (t-test)

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ccng1 WT standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 + MO4-tp53 standard conditions Prim-5 whole organism RTPCR
pmaip1 WT standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 + MO4-tp53 standard conditions Prim-5 whole organism RTPCR
tp53inp1 WT standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 standard conditions Prim-5 whole organism RTPCR
WT + MO3-nolc1 + MO4-tp53 standard conditions Prim-5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO3-nolc1 standard conditions Prim-5 apoptotic process increased occurrence, abnormal
Prim-5 head apoptotic, abnormal
Prim-5 whole organism ccng1 expression increased amount, abnormal
Prim-5 whole organism pmaip1 expression increased amount, abnormal
Prim-5 whole organism tp53inp1 expression increased amount, abnormal
Day 4 cranial cartilage morphology, abnormal
Day 4 embryonic cranial skeleton morphogenesis disrupted, abnormal
WT + MO3-nolc1 + MO4-tp53 standard conditions Prim-5 apoptotic process process quality, ameliorated
Prim-5 whole organism ccng1 expression increased amount, abnormal
Prim-5 whole organism pmaip1 expression amount, ameliorated
Prim-5 whole organism tp53inp1 expression amount, ameliorated
Day 4 cranial cartilage morphology, ameliorated
Day 4 embryonic cranial skeleton morphogenesis process quality, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Cell Death & Disease for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Death Dis.