(A to C) Cerebellar sections of (A) Tumor-bearing Ptch+/− and Ptch+/−/RESTTG mice (n=3) and (B) DAOY and DAOY-REST xenografts (n=3) and (C) human SHH subgroup patient derived xenografts (n=3) were analyzed by IHC for REST, PTCH1, and KI-67 expression using specific antibodies. (D)
PTCH1 and GLI1 mRNA expression profiles measured by microarray. Hierarchical clustering based on expression levels of neuronal differentiation markers divided the SHH MB patient samples into six distinct clusters. Each dot corresponds to one individual patient. (E)
Ptch1 and Gli1 mRNA expression was measured in WT (white bars) and RESTTG (gray bars) CGNPs after culturing with proliferation or differentiation media. WT data represents the mean ± S.D. from triplicate samples, RESTTG data represents two individual pups. Graph represents fold change compared to WT proliferating controls. For (A, B, and C), scale bars = 20 μm (40X). For (E), p values were calculated by paired two-tailed t test of ΔCp values: significance is indicated as not significant (ns), p<0.05 (*), p<0.01 (**), p<0.001 (***), or p<0.0001 (****).