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ZFIN ID: ZDB-FIG-121210-32
Wu et al., 2012 - Chemokine GPCR Signaling Inhibits β-Catenin during Zebrafish Axis Formation. PLoS Biology   10(10):e1001403 Full text @ PLoS Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Anatomical Terms:
Stage Range: 256-cell to Shield

Fig. 3

Ccr7 inhibits β-catenin activity via a Gsk3β-indepenedent mechanism.

(A) Hyper-dorsalized phenotypes caused by β-catenin overexpression (b,b2, 25 pg, n = 22/25), compared to control WT embryos (a), were suppressed by Ccr7 overexpression (c; 150 pg, n = 8/12). (d–f) Expansion of gsc expression domain in β-catenin overexpressing embryos (e), relative to control WT embryos (d), was suppressed by co-injection of ccr7 RNA (f). Animal views, dorsal to the right. (g) Frequency of embryos with gsc expression domain encompassing more (>180°) or less (<180°) than half of the embryo equator. (B) (a–c) LiCl-treated embryos (b; n = 16/20) show dorsalized phenotypes at 30 hpf compared to control embryos (a). LiCl-dependent dorsalization was suppressed by injection of ccr7 RNA (c; n = 8/20, two experiments). (d–f) gsc expression at shield stage (6 hpf) in control (d), LiCl-treated (e; n = 13/14), and LiCl-treated and ccr7 RNA-injected embryos (f; n = 9/12). Animal views, dorsal to the right. (g–i) β-catenin immunostaining at 256-cell stage in control (g), LiCl-treated (h, n = 9/10), and LiCl-treated embryos overexpressing Ccr7 (i; n = 9/11). Arrows point to a few β-catenin-positive nuclei in control embryos (g) and LiCl-treated embryos overexpressing Ccr7 (i). (C) Ccr7 antagonizes the ability of ΔNβ-catenin to rescue the ventralized ich mutant phenotype. (a) V1–V4 phenotypic classes, with V4 corresponding to the strongest ich phenotype. (b) Frequencies of the V1–V4 phenotypic classes of ich mutants injected with synthetic ΔNβ-catenin RNA alone or co-injected with ccr7 RNA. Injected amounts of RNAs in pg are shown below the graph, and the number of embryos in each group above each bar. (D) (a–c) Co-injections of ΔNβ-catenin-gfp RNA and MO1-ccr7 or ccr7 RNA showed that Ccr7 can downregulate β-catenin, shown at higher-magnification (d–f). Compared to control (a, d), ccr7 RNA overexpressing blastulae showed strongly decreased (b, e), while MO1-ccr7 injected blastulae showed increased, ΔNβ-catenin-GFP signal (c, f). H2B-RFP RNA was injected as nuclear background control (a2–c2 and higher magnification in d2–f2). (E) Western blot analysis of co-injection of ΔNβ-catenin-gfp RNA and ccr7 RNA or MO1-ccr7. Quantification of the relative protein level (signal intensity) from three independent immunoblots (bottom panel). * p<0.05.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
gsc WT standard conditions Shield shield ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-ctnnb IHC WT standard conditions 256-cell blastomere nucleus
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal PLoS Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Biol.