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ZIRC
ZFIN ID: ZDB-FIG-180126-35
Lee et al., 2017 - Functional validation of novel MKS3/TMEM67 mutations in COACH syndrome. Scientific Reports   7:10222 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Pec-fin
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Pec-fin

Fig. 4

Wnt signaling, but not Hh signaling, is reduced in tmem67 morphants. (a) Zebrafish embryos were injected with either control or tmem67 MOs [e2i2], subjected to WISH at 2.5 dpf with ptch1, ptch2 and gli1 riboprobes, and imaged under a light microscope. NS: not significant. Scale bar = 200 μm. (b) Total RNAs were extracted from WT or tmem67 morphant embryos at 2.5 dpf and subjected to quantitative real-time PCR with ptch1, ptch2, gli1 and axin2 primers. Expression levels of indicated mRNA were normalized to those of actin primers. NS: not significant. ** P < 0.01. (c) One-cell stage zebrafish embryos were injected with either control or tmem67 MOs [e2i2], probed with axin2 riboprobes at 2.5 dpf and imaged. For a rescue experiment, one-cell stage zebrafish embryos were sequentially injected with tmem67 MO [e2i2] and RNA encoding the indicated TMEM67 mutant, probed with axin2 riboprobes at 2.5 dpf and imaged. p.Tyr920* represents p.Tyr920ThrfsX40. Arrows indicate reduction of signals compared to those in control embryos. Scale bar = 200 μm. (d) Tg(Tcf/Lef-miniP:dGFP) embryos were injected with either control or tmem67 MOs [e3i3] and imaged at 1 dpf under a fluorescent microscope. Scale bar = 200 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
axin2 AB standard conditions Pec-fin brain ISH
Pec-fin whole organism RTPCR
AB + MO4-tmem67 standard conditions Pec-fin brain ISH
Pec-fin whole organism RTPCR
d2EGFP kyu1Tg standard conditions Prim-5 brain IFL
kyu1Tg + MO4-tmem67 standard conditions Prim-5 brain IFL
gli1 AB standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
AB + MO4-tmem67 standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
ptch1 AB standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
AB + MO4-tmem67 standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
ptch2 AB standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
AB + MO4-tmem67 standard conditions Pec-fin head ISH
Pec-fin whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO4-tmem67 standard conditions Prim-5 Wnt signaling pathway disrupted, abnormal
Pec-fin brain axin2 expression decreased amount, abnormal
Pec-fin whole organism axin2 expression decreased amount, abnormal
kyu1Tg + MO4-tmem67 standard conditions Prim-5 brain d2EGFP expression decreased amount, abnormal
Prim-5 Wnt signaling pathway disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Scientific Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Sci. Rep.