Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information | ||||
|---|---|---|---|---|---|---|---|---|
| Gene (Homo sapiens) | HES3 | NA | NM_001024598 | |||||
| Gene (Homo sapiens) | PAX3-FOXO1 | PMID:8275086; PMID:8221646 |
NM_181457; NM_002015.3 |
exons 1–7 of NM_181457 fused to exons 2–3 of NM_002015.3 |
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| Gene (Mus musculus) | Pax3 | NA | NM_008781 | |||||
| Gene (Danio rerio) | her3 | NA | NM_131080 | |||||
| Strain, strain background (Danio rerio AB) |
AB Wildtype | Zebrafish International Resource Center (ZIRC) |
ZIRC:ZL1 | https://zebrafish.org/fish/lineAll.php | ||||
| Strain, strain background (Danio rerio AB/TL) |
AB\TL Wildtype | NA | Cross of AB and TL | |||||
| Strain, strain background (Danio rerio TL) |
TL Wildtype | ZIRC | ZIRC:ZL86 | https://zebrafish.org/fish/lineAll.php | ||||
| Strain, strain background (Danio rerio WIK) |
WIK Wildtype | ZIRC | ZIRC:ZL84 | https://zebrafish.org/fish/lineAll.php | ||||
| Strain, strain background (Danio rerio tp53M214K mutant) |
tp53M214K mutant | PMID:15630097; available from ZIRC |
ZIRC:ZL1057 | https://zebrafish.org/fish/lineAll.php | ||||
| Genetic reagent | Tol2 transposase mRNA | PMID:16959904 | Injected at 50 ng/uL | |||||
| Cell line (Mus musculus) |
C2C12 | ATCC | ATCC:CRL-1772 | Maintained in DMEM + 10% FBS+1X Antimycotic-Antibiotic |
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| Cell line (Homo sapiens) |
Rh30 | ATCC | ATCC:CRL-2061 | Maintained in RPMI-1640 + 10% FBS+1X Antimycotic-Antibiotic |
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| Transfected construct (Homo sapiens) |
CMV-HES3 (MYC and FLAG tagged) |
Origene | Origene:RC224630 | |||||
| Transfected construct (NA) |
CMV-Empty | this paper |
HES3 excised using EcoR1 and Mlu1 and re-ligated |
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| Antibody | MF20 (mouse monoclonal) | Developmental Studies Hybridoma Bank |
DSHB:MF20c | 1:40 in cells, 1:100 in zebrafish embryo whole mounts, 1:1000 Western blot |
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| Antibody | alpha-Tubulin DM1A (mouse monoclonal) |
Cell Signaling | Cell Signaling:3873 | 1:1000 dilution | ||||
| Antibody | MyoD 5.8A (mouse monoclonal) |
Thermo | Thermo:MA5-12902 | 1:1000 dilution | ||||
| Antibody | MYC 71D10 (rabbit monoclonal) |
Cell Signaling | Cell Signaling:2278 | 1:1000 dilution | ||||
| Antibody | GFP (rabbit polyclonal) | MBL International Corporation |
MBL:598 | 1:1000 dilution | ||||
| Antibody | GFP-488 (rabbit polyclonal) |
Thermo | Thermo:A-21311 | 1:500 dilution | ||||
| Antibody | Alexa 488 or 594 secondaries |
Thermo | 1:500 dilution | |||||
| Antibody | HRP conjugate secondaries |
BioRad | 1:20000 dilution | |||||
| Other | DAPI stain (ProLong Gold Antifade mounting media with DAPI) |
Thermo | Thermo:P36931 | |||||
| Recombinant DNA reagent |
p5E beta actin | PMID:17937395 | ||||||
| Recombinant DNA reagent |
p5E cmv | PMID:17937395 | ||||||
| Recombinant DNA reagent |
p5E mcs | PMID:17937395 | ||||||
| Recombinant DNA reagent |
p5E ubi | PMID:21138979; available from Addgene |
Addgene:27320 | |||||
| Recombinant DNA reagent |
p5E unc503 | PMID:23444339; available from Addgene |
Addgene:64020 | |||||
| Recombinant DNA reagent |
p5E fli1a | PMID:17948311; available from Addgene |
Addgene:31160 | |||||
| Recombinant DNA reagent |
p5E mitfa | James Lister; available from Addgene |
Addgene:81234 | |||||
| Recombinant DNA reagent |
pmE beta globin splice acceptor |
PMID:15239961 | ||||||
| Recombinant DNA reagent |
pmE GFP2A | PMID:17941043 | ||||||
| Recombinant DNA reagent |
pmE mCherry2A | PMID:17941043 | ||||||
| Recombinant DNA reagent |
p3E PAX3-FOXO1 | this paper | attb2r/attb3 sites added with primers in Supplementary file 3 by high-fidelity PCR |
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| Recombinant DNA reagent |
p3E Pax3 | this paper | attb2r/attb3 sites added with primers in Supplementary file 3 by high-fidelity PCR |
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| Recombinant DNA reagent |
p3E HES3 | this paper | attb2r/attb3 sites added with primers in Supplementary file 3 by high-fidelity PCR |
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| Recombinant DNA reagent |
p3E SV40 late poly A | PMID:17937395 | ||||||
| Recombinant DNA reagent |
p3E 2A-mCherry | PMID:23462469; available from Addgene |
Addgene:26031 | |||||
| Recombinant DNA reagent |
pDONRP2R-P3 (3′ donor vector; attP2R-P3 flanking chlor/ccdB cassette) |
Invitrogen | Invitrogen:pDONR P2R-P3 |
Used to generate p3E's from this paper |
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| Recombinant DNA reagent |
pDestTol2pA2 destination vector |
PMID:17937395 | ||||||
| Recombinant DNA reagent |
BetaActin-GFP2A-pA | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
BetaActin-mCherry2A-pA | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
BetaActin-GFP2A- 2AmCherry |
this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
BetaActin-mCherry2A-Pax3 | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
BetaActin-GFP2A-Pax3 | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
BetaActin-mCherry2A-HES3 | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
BetaActin-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
CMV-GFP2A-pA | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
CMV-mCherry2A-pA | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
CMV-GFP2A-Pax3 | this paper | Generated by Gateway Cloning |
|||||
| Recombinant DNA reagent |
CMV-mCherry2A-HES3 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
CMV-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
ubi-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
mitfa-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
fli1-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
unc503-GFP2A-PAX3FOXO1 | this paper | Generated by Gateway Cloning |
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| Recombinant DNA reagent |
SpliceAcceptor- GFP2A-PAX3FOXO1 |
this paper | Generated by Gateway Cloning |
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| Sequence-based reagent |
GFP Morpholino | Gene Tools | Gene Tools: GFP Morpholino |
(5' ACAGCTCCTCGCCC TTGCTCACCAT 3') |
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| Commercial assay or kit |
ApopTag Red In Situ Apoptosis Detection Kit |
Millipore | Millipore:S7165 | |||||
| Commercial assay or kit |
Affymetrix Zebrafish Gene 1.1 ST Array strip |
Affymetrix | Affymetrix:901802 | |||||
| Commercial assay or kit (Mus musculus) |
M384 Rhabdomy osarcoma 384 well panel |
BioRad | BioRad:M384 Rhabdomyosarcoma |
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| Commercial assay or kit (Homo sapiens) |
H384 Tumor Metastasis (SAB Target List) 384 well panel |
BioRad | BioRad:H384 Tumor Metastasis (SAB Target List) |
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| Chemical compound, drug |
Geneticin (G418) | Thermo | Thermo:10131027 | Select at 1 mg/mL | ||||
| Software, algorithm | ImageJ | http://imageJ.nih.gov/ij | ||||||
| Software, algorithm | GraphPad Prism 7.0 c | https://www.graphpad.com | ||||||
| Software, algorithm | Rv3.3.1 | https://www.R-project.org | ||||||