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Focus formation (NIH-3T3) assay

SZ Sofia Zanotti
SV Suzanne Vanhauwaert
CN Christophe Van Neste
VO Volodimir Olexiouk
JL Jolien Van Laere
MV Marlies Verschuuren
JM Joni Van der Meulen
LM Liselot M. Mus
KD Kaat Durinck
LT Laurentijn Tilleman
DD Dieter Deforce
FN Filip Van Nieuwerburgh
MH Michael D. Hogarty
BD Bieke Decaesteker
WV Winnok H. De Vos
FS Frank Speleman
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To quantify the impact of the oncogene MYCN on the proliferation rate over time and several passages, a NIH-3T3-assay was applied as previously described by Todaro et al.37. In brief, cells with an inducible MYCN OFF/ON system, respectively either maintained in control conditions (EtOH) or induced MYCN (4-OHT), were plated in T25 flasks at a density of 0,15 million cells. The cumulative increase in cell number was calculated according to the formula log(Nf/Ni)/log2, where Ni and Nf are the initial and final numbers of cells plated and counted after 3 or 4 days respectively.

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