Cell lines RH30 and RH4 correspond to aRMS with PAX3/FOXO1 translocation, RH18 to a translocation-negative aRMS and RD and HTB82 to eRMS subtype. Some authors have attributed the HTB82 cell line to a rhabdoid tumour since it bears a mutation in the SMARCB1 gene (Khan et al, 2001 (link); Hinson et al, 2013 (link)). All RMS cell lines were previously authenticated by STR-based DNA profiling, grown in MEM media (Biowest, Barcelona, Spain), supplemented with 10% foetal bovine serum (Sigma-Aldrich, Madrid, Spain), 2 mM L-glutamine, 1 mM sodium pyruvate and 1 × non-essential amino acids (all reagents from Biowest). All cell lines were obtained from American Type Culture Collection, except RH18 and RD cell lines which were a kind gift from Dr Beat Schäfer. The SMO inhibitor Vismodegib (GDC-0449) was purchased from Selleckchem (Madrid, Spain). The antibody MEDI-5304 was kindly provided by MedImmune (Cambridge, UK).
Vismodegib gdc 0449
Manufactured by Selleck Chemicals
Sourced in Spain
Vismodegib (GDC-0449) is a small molecule inhibitor of the Hedgehog signaling pathway. It is used as a research tool to study the Hedgehog signaling pathway in various experimental models.
Automatically generated - may contain errors
Lab products found in correlation
Sodium pyruvate, by Biowest (1 mentions)
Foetal bovine serum (fbs), by Merck Group (1 mentions)
Mem media, by Biowest (1 mentions)
L glutamine, by Biowest (1 mentions)
Non essential amino acids (neaa), by Biowest (1 mentions)
Torin1, by Bio-Techne (1 mentions)
Z vad fmk fmk001, by R&D Systems (1 mentions)
3 2 4 dichloro 5 methoxyphenyl 2 3 dihydro 2 thioxo 4 1h quinazolinone mdivi 1 bml cm127, by Enzo Life Sciences (1 mentions)
Ciliobrevin a1, by Bio-Techne (1 mentions)
N acetylcysteine nac a9165, by Merck Group (1 mentions)
Ccg 1423, by Cayman Chemical (1 mentions)
Celltiter 96 aqueous one solution, by Promega (1 mentions)
Dasatinib, by Selleck Chemicals (1 mentions)
Gant61, by Selleck Chemicals (1 mentions)
Ly294002 hydrochloride, by Bio-Techne (1 mentions)
Hpi 1, by Bio-Techne (1 mentions)
U0126, by Bio-Techne (1 mentions)
Flavopiridol, by Selleck Chemicals (1 mentions)
Ruxolitinib, by Selleck Chemicals (1 mentions)
1 azakenpaullone, by Selleck Chemicals (1 mentions)
5 protocols using vismodegib gdc 0449
1
Rhabdomyosarcoma Cell Line Characterization
2
Cellular Metabolism Modulation Compounds
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Bafilomycin A1 (B1793), Carbonyl cyanide m-chlorophenyl hydrazine (CCCP, C2759), rotenone (R8875), 1-methyl-4-phenylpyridinium (MPP+, D048), doxycycline (D9891), and N-acetyl-cysteine (NAC, A9165) were purchased from Sigma-Aldrich (St. Louis, MO). 3-(2,4-dichloro-5-methoxyphenyl)-2,3-dihydro-2-thioxo-4(1H)-quinazolinone (Mdivi-1, BML-CM127) was purchased from Enzo Life Sciences (Farmingdale, NY). Ciliobrevin A1 (#4529) and Torin-1 (#4247) were purchased from Tocris Bioscience (Bristol, UK). Vismodegib (GDC-0449) was purchased from Selleckchem (Munich, Germany). zVAD-FMK (FMK001) was purchased from R&D systems (Minneapolis, MN). 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, HY-15608) was obtained from MedChem Express (Monmouth Junction, NJ).
3
Evaluating Compound Cytotoxicity in Ras-Activated Cells
4
Evaluation of Hedgehog Pathway Inhibitors
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The tyrosine kinase inhibitor (TKI) dasatinib, SMO agonist (SAG)- hydrochloride, GLI1/2 inhibitor GANT61, and SMO antagonist vismodegib (GDC-0449) were purchased from Selleckchem (Houston, TX). GLI1/2 inhibitor HPI-1, PI3Kα/δ/β inhibitor LY-294002 hydrochloride, and MEK1/2 inhibitor U0126 were purchased from Tocris (Minneapolis, MN). High activity recombinant Human Sonic Hedgehog (SHH) Protein was purchased from R&D Systems (Minneapolis, MN) and arsenic trioxide (A2O3 or ATO) from Millipore Sigma (Burlington, MA).
5
Transcriptional Inhibitor Screening in Halocynthia Embryos
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Actinomycin D (Sigma) was diluted into DMSO at 10 mg/mL stock. The stock solution was diluted into ASW to a final concentration of 40 μg/mL. This concentration was reported to block transcription in Halocynthia embryos 42 . Flavopiridol (Selleck chemicals) was diluted into water to 10mM stock. The stock solution was diluted into ASW to final concentration 1 and 10 μM. The transcriptional inhibitor treated embryos were fixed by MEM-PFA (4% PFA, 0.1M MOPS, 0.5M NaCl, 1mM EGTA, 2mM MgSO 4 ) after 1 hour inhibitor treatment, and used for in situ hybridization.
1-Azakenpaullone (S7193; Selleckchem; 43 ), Ruxolitinib (INCB018424; S1378; Selleckchem), Vismodegib (GDC-0449; S1082; Selleckchem), DAPT (Millipore Sigma), SB431542 (S1067; Selleckchem; 35 ), U0126 (Cell Signaling Technology;
) and Dorsomorphin (Selleckchem; 35, 43 ) were used to perturb define signaling pathways as described in corresponding references. These treatments were done in a final concentration of 10 μM for 2 or 4 hours. The inhibitor-treated embryos were fixed by MEM-PFA after 2 hours inhibitor treatment, and used for in situ hybridization.
1-Azakenpaullone (S7193; Selleckchem; 43 ), Ruxolitinib (INCB018424; S1378; Selleckchem), Vismodegib (GDC-0449; S1082; Selleckchem), DAPT (Millipore Sigma), SB431542 (S1067; Selleckchem; 35 ), U0126 (Cell Signaling Technology;
) and Dorsomorphin (Selleckchem; 35, 43 ) were used to perturb define signaling pathways as described in corresponding references. These treatments were done in a final concentration of 10 μM for 2 or 4 hours. The inhibitor-treated embryos were fixed by MEM-PFA after 2 hours inhibitor treatment, and used for in situ hybridization.
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