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ZFIN ID: ZDB-FIG-140325-64
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:
Genes:
Antibodies:
Fish:
Condition:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-15
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: 14-19 somites to Prim-15

Fig. 4

Regulation of Gli processing and activity by Kif7.

(A) Western blot analysis showing Gli2a processing in wild-type (WT), MZkif7 (MZ) and Shh mRNA injected (Shh) embryos at 18 hpf (left panel) and at 22 hpf (right panel) compared to the same set treated with cyclopamine (CycA). Full-length (FL) and repressor (R) forms of Gli2a are indicated. The lower panels shows the same blot re-probed with Kif7 and γ-tubulin (loading control) antibodies (B,C) Quantification of the Gli2a FL:R ratio normalized to WT at 18 hpf (B) and 22 hpf (C). Bar graphs numbered 1–6 represent lanes on the blots in (A) from left to right. Error bars represent standard deviation obtained from three independent Western blots including those shown in (A). Single asterisk: P<0.02; double asterisk: P<0.001. (D–H) Parasagittal optical sections of 30 hpf embryos of different genotypes showing expression of Eng-expressing muscle cells as revealed by mAb4D9 (green). The MZkif7 phenotype (D) is largely unaffected by removal of Gli2a (F), other than the loss of the some MP cells (arrows); by contrast, Eng expression is nearly eliminated in the MZkif7;gli1 double mutant embryo (G), despite Gli1 being dispensable for Eng expression in the presence of Kif7 function (E). Knockdown of gli2b and gli3 activity using morpholinos in gli1 mutants (H) has no effect on Eng expression in the myotome. Scale bar: 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
gli2a Ab1-gli2a WT standard conditions 14-19 somites whole organism WB
Ab1-gli2a 26+ somites whole organism WB
Ab1-gli2a WT chemical treatment: Cyclopamine 14-19 somites whole organism WB
Ab1-gli2a 26+ somites whole organism WB
Ab1-gli2a kif7i271/i271 standard conditions 14-19 somites whole organism WB
Ab1-gli2a 26+ somites whole organism WB
Ab1-gli2a kif7i271/i271 chemical treatment: Cyclopamine 14-19 somites whole organism WB
Ab1-gli2a 26+ somites whole organism WB
kif7 Ab1-kif7 WT standard conditions 14-19 somites whole organism WB
Ab1-kif7 26+ somites whole organism WB
Ab1-kif7 WT chemical treatment: Cyclopamine 14-19 somites whole organism WB
Ab1-kif7 26+ somites whole organism WB
Ab1-kif7 kif7i271/i271 standard conditions 14-19 somites Not Detected whole organism WB
Ab1-kif7 26+ somites Not Detected whole organism WB
Ab1-kif7 kif7i271/i271 chemical treatment: Cyclopamine 14-19 somites Not Detected whole organism WB
Ab1-kif7 26+ somites Not Detected whole organism WB
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-eng IHC gli1ts269/ts269 standard conditions Prim-15 fast muscle cell
IHC Prim-15 muscle pioneer
IHC gli1ts269/ts269 ; kif7i271/i271 standard conditions Prim-15 muscle pioneer
IHC Prim-15 myotome
IHC gli2aty119/ty119 ; kif7i271/i271 standard conditions Prim-15 muscle pioneer
IHC Prim-15 myotome
IHC kif7i271/i271 standard conditions Prim-15 fast muscle cell
IHC Prim-15 muscle pioneer
IHC Prim-15 myotome
Phenotype Details
Fish Conditions Stage Phenotype
gli1ts269/ts269 ; kif7i271/i271 standard conditions Prim-15 muscle pioneer decreased amount, abnormal
Prim-15 muscle pioneer ab1-eng labeling decreased distribution, abnormal
Prim-15 myotome ab1-eng labeling decreased distribution, abnormal
gli2aty119/ty119 ; kif7i271/i271 standard conditions Prim-15 muscle pioneer decreased amount, abnormal
Prim-15 myotome ab1-eng labeling decreased distribution, abnormal
kif7i271/i271 standard conditions 14-19 somites regulation of protein processing process quality, abnormal
14-19 somites whole organism kif7 expression absent, abnormal
14-19 somites whole organism gli2a expression decreased amount, abnormal
26+ somites whole organism kif7 expression absent, abnormal
Prim-15 myotome ab1-eng labeling increased distribution, abnormal
kif7i271/i271 chemical treatment: Cyclopamine 14-19 somites whole organism kif7 expression absent, abnormal
14-19 somites whole organism gli2a expression decreased amount, abnormal
26+ somites whole organism kif7 expression absent, 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.