ZFIN ID: ZDB-FIG-191230-450
Tuttle et al., 2019 - Retrograde Ret signaling controls sensory pioneer axon outgrowth. eLIFE   8 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage: Prim-15
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-15

Figure 7 Assessment of anti-Myo10 antibody and <italic>myo10l1</italic> gRNA efficiency.

(A) Myo10 antibody staining for neurons in embryos injected with myo10l1-eGFP BAC. Rohon-Beard neurons expressing Myo10l1-eGFP (A) are also positive for Myo10 antibody staining (A’). (B) Scale diagram of zebrafish Myo10l1 protein. gRNAs target distinct sites in myo10l1 exons corresponding to specific regions of the protein (red arrowheads). PH = Pleckstrin homology domain, MyTH = Myosin tail homology four domain, FERM = 4.1 protein, Ezrin, Radixin, Moesin domain. (C) Amplification of targeted CRISPR cut target regions containing restriction cut sites. Gel electrophoresis (2% agarose gel) of PCR amplification of each CRISPR cut site from genomic DNA of individual four dpf larvae injected with gRNA triplex cocktail that display significant pLLG truncation or uninjected embryos. Following PCR, amplicons are incubated with restriction enzymes that cut near CRISPR target sites. Uninjected embryos have near complete digest of amplicons at each CRISPR target site, but triplex-injected crispants that display pLLG truncation show substantial amounts of resistance to digest. *=1 kb plus ladder.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP nl1Tg standard conditions Prim-15 posterior lateral line ganglion IFL
Prim-15 posterior lateral line ganglion growth cone IFL
rethu2846/hu2846; nl1Tg standard conditions Prim-15 posterior lateral line ganglion IFL
Prim-15 posterior lateral line ganglion growth cone IFL
myo10l1 nl1Tg standard conditions Prim-15 posterior lateral line ganglion ISH
rethu2846/hu2846; nl1Tg standard conditions Prim-15 posterior lateral line ganglion ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-myo10 IHC mapk8ip3nl7/nl7; nl1Tg standard conditions Prim-15 posterior lateral line ganglion growth cone
IHC nl1Tg standard conditions Prim-15 posterior lateral line ganglion growth cone
IHC rethu2846/hu2846; nl1Tg standard conditions Prim-15 posterior lateral line ganglion growth cone
Phenotype Details
Fish Conditions Stage Phenotype
mapk8ip3nl7/nl7; nl1Tg standard conditions Prim-15 posterior lateral line ganglion growth cone Ab1-myo10 labeling decreased amount, abnormal
nl1Tg + CRISPR1-myo10l1 + CRISPR2-myo10l1 + CRISPR3-myo10l1 standard conditions Prim-15 posterior lateral line ganglion axon truncated, abnormal
rethu2846/hu2846; nl1Tg standard conditions Prim-15 posterior lateral line ganglion myo10l1 expression decreased amount, abnormal
Prim-15 posterior lateral line ganglion axon truncated, abnormal
Prim-15 posterior lateral line ganglion growth cone Ab1-myo10 labeling decreased amount, abnormal
rethu2846/hu2846; nl1Tg + CRISPR1-myo10l1 + CRISPR2-myo10l1 + CRISPR3-myo10l1 standard conditions Prim-15 posterior lateral line ganglion axon truncated, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife