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ZFIN ID: ZDB-FIG-180626-14
Mönnich et al., 2018 - CEP128 Localizes to the Subdistal Appendages of the Mother Centriole and Regulates TGF-β/BMP Signaling at the Primary Cilium. Cell Reports   22:2584-2592 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Shield to 20-25 somites
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Shield to 20-25 somites

Fig. 2

CEP128 Regulates TGF-β/BMP Signaling in Zebrafish

(A) Phenotype of cep128-depleted embryos at the 10-somite stage. Embryos show mild (note the pointy tail of the embryo, arrow) to severe (egg-shaped yolk sac) dorsalized morphology. Lateral views, anterior to left.

(B) Numbers for the phenotypic analysis in (A). Two to four independent experiments.

(C–E) Dorsoventral patterning genes are misexpressed in cep128 splx morphants. Areas of expression of dorsal genes chd (C) and gsc (D) are increased in morphants; the ventrally expressed gene eve1 (E) is slightly decreased. Animal pole views, dorsal to right.

(F) The phosphorylation level of Smad1/5/8 is slightly reduced in morphants. The thickened embryonic tissue in the morphants indicates dorsalization of the embryo (arrow). Lateral views, dorsal to right.

(G) myoD1 expression (blue) shows thin and elongated somites in morphants, with adaxial cells extending around the tail bud (arrowhead); the notochord (ta expression, red) is thickened and bent. Dorsal views, anterior to top.

(H) ta expression is ectopic and unstructured in the notochord and tailbud areas in cep128 morphants.

(I–L) WB analysis of embryos at 75% epiboly (I and K) and quantification of relative band intensities (J and L) show that phosphorylation levels of Smad1/5/8 and Smad2 are reduced significantly in morphants compared to WT. Graphs show means ± SDs (n = 3). ∗∗p ≤ 0.01; ∗∗∗∗p ≤ 0.0001.

See also Figure S3.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
chrd AB/TL standard conditions Shield shield ISH
AB/TL + MO1-cep128 standard conditions Shield shield ISH
eve1 AB/TL standard conditions 75%-epiboly margin ISH
AB/TL + MO1-cep128 standard conditions 75%-epiboly margin ISH
gsc AB/TL standard conditions Shield shield ISH
AB/TL + MO1-cep128 standard conditions Shield shield ISH
tbxta AB/TL standard conditions 20-25 somites notochord ISH
20-25 somites tail bud ISH
AB/TL + MO1-cep128 standard conditions 20-25 somites notochord ISH
20-25 somites tail bud ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab11-smad2 WB AB/TL standard conditions 75%-epiboly whole organism
WB AB/TL + MO1-cep128 standard conditions 75%-epiboly whole organism
Ab12-smad WB AB/TL standard conditions 75%-epiboly whole organism
WB AB/TL + MO1-cep128 standard conditions 75%-epiboly whole organism
Phenotype Details
Fish Conditions Stage Phenotype
AB/TL + MO1-cep128 standard conditions Shield shield chrd expression increased distribution, abnormal
Shield shield gsc expression increased distribution, abnormal
75%-epiboly margin eve1 expression decreased amount, abnormal
75%-epiboly whole organism Ab11-smad2 labeling decreased amount, abnormal
75%-epiboly whole organism Ab12-smad labeling decreased amount, abnormal
10-13 somites whole organism dorsalized, abnormal
10-13 somites whole organism ovate, abnormal
14-19 somites notochord bent, abnormal
14-19 somites notochord increased thickness, abnormal
14-19 somites somite decreased thickness, abnormal
14-19 somites somite elongated, abnormal
20-25 somites notochord tbxta expression spatial pattern, abnormal
20-25 somites tail bud tbxta expression spatial pattern, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.