Molecular subtypes of medulloblastomas were detected, as recommended by the 2016 WHO classification:
WNT-activated tumors were identified by the presence of at least two features: CTNNB1 mutation, immunohistochemical positive nuclear reaction against β-catenin (#760-4242, clone 14, Cell Margue, dilution 1.68 µg/ml) and the presence of chromosome 6 monosomy detected by multiplex ligation-dependent probe amplification (MLPA).
SHH-activated tumors were determined by the presence of immunohistochemical positive reaction with anti-GAB1 (Abcam, Cambridge, USA, #ab27439 and/or ab #59362, dilution 1:100) and anti-YAP1 (Santa Cruz Biotechnology, Dallas, USA, #sc-101199, dilution 1:50) antibodies, as described by Ellison et al. [12 (link)].
The remaining tumors that were negative for the above features were assigned as non-WNT, non-SHH tumors.
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