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Material and Methods

We are happy to share all materials and methods described in our publications. Please do not hesitate to contact us in this regard. Furthermore, we here want to briefly list the methods available in our work group, and additional information concerning the materials provided by us.

Methods (selected)

Cell culture: cell culture of neuroblastoma, retinoblastoma and medulloblastoma cell lines; cell proliferation and apoptosis assays; invasion-( Boyden-chamber assay); transfection by electroporation and lipofection; Tet-inducible gene expression; siRNA and shRNA

DNA-Repair: H2AX assay; NHEJ-religation assay

Flow Cytometry (Beckmann Coulter Cytomics 500): cell cycle analysis

Murine models: ectopic (s.c.) mouse models of medulloblastoma and neuroblastoma; orthotopic murine model; use of Tet-inducible cell lines in vivo; syngenic murine neuroblastoma models (C1300 / NXS2 cell lines in A/J Mäusen, own A/J colony); syngenic transplantation of TH-NMYC cell lines into 129/SvJ mice; Ivis Lumina in vivo imaging; application of substances i.p., i.v. (tail vain) and by oral gavage; blood sampling; breeding and genotyping of transgenic mice; breeding and analysis of the TH-NMYC mouse model (Weiss et al., EMBO 1997); breeding and analysis of the SMOA1 and PTCH+/- medulloblastoma mouse models

Histology: parafin and cryo sections; various IHC stainings; LacZ staining

Material (selected)

SY5Y-TR-TrkA: Neuroblastoma cell line for Tet-inducible expression of the TrkA neurotrophin receptor. This cell line is provided by our group within the FP7 EU project ASSET. A handbook for this cell lines is available to be downloaded.

SHEP-TR-miR-17-92: Neuroblastoma cell line for Tet-inducible expression of the miR-17-92 cluster.

Tet-repressor transfected neuroblastoma cell lines: We transfected several neuroblastoma cell lines with the Tet-repressor by electroporation (Tet-on-system provide by Invitrogen, pcDNA6/TR), and subsequently raised single cell clones by limited dilution. The single cell clones were subsequently chracterized and tested. We are happy to provide the following cell lines: NLF-TR, CLBGA-TR, SKNAS-TR, IMR32-TR and SY5Y-TR.