header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-140325-68
Maurya et al., 2013 - Positive and negative regulation of Gli activity by Kif7 in the zebrafish embryo. PLoS Genetics   9(12):e1003955 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Prim-15
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-15

Fig. 6

Kif7 modulates Gli2a localization and its association with Sufu.

(A) Western blot analysis of anti-Gli2a immune-precipitates of WT and MZkif7 (MZ) embryos, showing association of Kif7 with Gli2a in wild-type embryos and increased association of Sufu with Gli2a in MZkif7 embryos, as indicated by the ratio of the normalized intensities of the MZ to WT signals. (B) Western blot analysis of anti-Sufu immuno-precipitates of WT, Shh RNA injected (Shh), cyclopamine (cycA) exposed and MZkif7 (MZ) embryos; the Gli2aFL:Sufu ratios normalized to wild-type are shown below each lane. Error bars represent standard deviation obtained from three independent biological replicates. Note the increase in levels of Gli2aFL that co-precipitates with Sufu in MZkif7 embryos. (C,D) Localization of a functional eGFP tagged Gli2a protein (green) to primary cilia in wild-type (C) and MZkif7 (D) embryos. The axonemes of the primary cilia are marked by acetylated α-tubulin (red) and the basal bodies by γ-tubulin (blue) staining. In wild-type, Gli2a localizes to the tip of the cilia whereas in MZkif7, localization is restricted to the base of the cilia; the lower two panels in (D) are the same images as in the upper panels but with the red channel removed to show the juxtaposed GFP-Gli2a and γ-tubulin signals more clearly. Scale bar: 5 μm. (E) Western blot of wild-type (WT) and sufu morpholino-injected (sufu MO) embryo extracts probed with anti-Sufu and anti γ-tubulin, showing significant depletion of Sufu levels in the morphants relative to wild-type (MO/WT). (F–K) Parasagittal optical sections of 30 hpf wild-type (WT) or MZkif7 embryos showing the effect on eng2a:eGFP expression of morpholino mediated knockdown of gli1 (H,I) or gli1 and sufu (J,K). Depletion of gli1 in WT embryos (H) has no discernible effect, whereas it causes a drastic suppression of the ectopic expression in MZkif7 (I). This suppression is abrogated by simultaneous removal of Sufu and Gli1 from MZkif7 embryos (K). Scale bar: 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP i233Tg standard conditions Prim-15 fast muscle cell IFL
i233Tg + MO1-gli1 standard conditions Prim-15 fast muscle cell IFL
Prim-15 myotome IFL
i233Tg + MO1-gli1 + MO1-sufu standard conditions Prim-15 fast muscle cell IFL
Prim-15 myotome IFL
kif7i271/i271; i233Tg standard conditions Prim-15 fast muscle cell IFL
Prim-15 myotome IFL
kif7i271/i271; i233Tg + MO1-gli1 standard conditions Prim-15 fast muscle cell IFL
Prim-15 myotome IFL
kif7i271/i271; i233Tg + MO1-gli1 + MO1-sufu standard conditions Prim-15 fast muscle cell IFL
Prim-15 myotome IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
kif7i271/i271; i233Tg standard conditions Prim-15 myotome EGFP expression increased distribution, abnormal
kif7i271/i271; i233Tg + MO1-gli1 standard conditions Prim-15 myotome EGFP expression decreased distribution, abnormal
kif7i271/i271; i233Tg + MO1-gli1 + MO1-sufu standard conditions Prim-15 myotome EGFP expression increased distribution, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.