Failure of human rhombic lip differentiation constitutes medulloblastoma

Single-nucleus RNA sequencing was performed on Group 3 medulloblastoma tumorspheres expresssing high vs. low levels of OTX2 OTX2 levels were knocked down in HDMB03 and MB3W1 Group 3 MB cells (2X105 cells/well) as previously described (Zagozewski et al., Nature Comm, 2020; Stromecki et al., Molecular Oncology, 2018). Briefly, OTX2 was silenced using Silencer Select siRNAs 9931 or 9932 (Life Technologies) at a concentration of 30 nM. A non-silencing (scramble) served as the negative control. Samples were collected at 72 hours for RNA extraction and sequencing.