Generation of Orthotopic MB-G3 Mouse Models

To develop EP-based MB_G3 models, in utero EP was performed.^{37 (link)} Only the animals showing luciferase signal by P7 were selected for further analysis. Tumor growth was measured every 1–2 week by bioluminescence imaging of luciferase activity using a Xenogen IVIS system (PerkinElmer, Waltham, MA, USA).^{41 (link)} Orthotopic MB_G3 mouse models were generated by cranial implants of purified GNPs from [Trp53^−/−; Cdkn2c^−/−], [Trp53^Fl/-; Atoh1-CreER], [Trp53^Fl/-; Prom1-CreER] and [Trp53^Fl/-] P6–7 pups, infected with retroviruses carrying Myc and red fluorescent protein according to the previous study.^{16 (link)} No specific randomization or blinding was performed. Tumor cell purification and genomic DNA and total RNA extraction for genotyping and Affymetrix microarray analysis were described previously.^{16 (link)} For detection of non-recombined and recombined alleles of Trp53 in Atoh1ER-MYC and Prom1ER-MYC MBs, 1F/1R (370 bp) and 1F/10R (612 bp) primer sets were used, respectively.^{42 (link)} For internal control, endogenous Prom1 (563 bp)^{43 (link)} and Ptch1 (220 bp)^{17 (link)} were detected. Comparison of survival curves was performed by calculation of two-tailed P-value using the GraphPad Prism software (GraphPad Software, Inc., La Jolla, CA, USA).

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Kawauchi D., Ogg R.J., Liu L., Shih D.J., Finkelstein D., Murphy B.L., Rehg J.E., Korshunov A., Calabrese C., Zindy F., Phoenix T., Kawaguchi Y., Gronych J., Gilbertson R.J., Lichter P., Gajjar A., Kool M., Northcott P.A., Pfister S.M, & Roussel M.F. (2017). Novel MYC-driven medulloblastoma models from multiple embryonic cerebellar cells. Oncogene, 36(37), 5231-5242.

Publication 2017

Xenogen IVIS system GraphPad Prism software

Alleles Animals Cdkn2c Cell Cranial Genomic Luciferase Microarray analysis Mouse Primer Prism Ptch1 Red fluorescent protein Retroviruses Trp53 Tumor Utero

Corresponding Organization : St. Jude Children's Research Hospital

Other organizations : Hospital for Sick Children, German Cancer Research Center, Kyoto University, University Hospital Heidelberg

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Variable analysis

independent variables

Genetic manipulations (Trp53, Cdkn2c, Atoh1-CreER, Prom1-CreER)
Viral infections (Myc and red fluorescent protein retroviruses)

dependent variables

Luciferase signal by P7
Tumor growth measured by bioluminescence imaging

control variables

No specific randomization or blinding was performed

controls

Positive control: Luciferase signal by P7 used for selecting animals for further analysis
Negative control: Not explicitly mentioned

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