Supplemental figure and table legends
Supplemental Figure S1. Snf5 inactivation causes neurodegeneration
(A-D) H&E (A), MBP (B), Olig 2 (C), Beta III Tubulin (D) of sagittal brain sections of the hippocampal region of an adult control mouse (left top and bottom) and Snf5F/FGFAP-Cre mouse (right top and bottom), demonstrating the granular dentate gyrus (DG) and the CA1, CA2, and CA3 areas (top). The corpus callosum (cc) and the cerebral cortex (on top of the cc) are also shown. In comparison to the control (left), the Snf5F/FGFAP-Cre mouse (right) demonstrates prominent vacuolization with disruption of the corpus callosum. At higher magnification (bottom), the pyramidal neurons of the CA1 area of the hippocampus are mostly well preserved (A). The profound loss of axon and myelin of the cc is reflected in the reduced stain of MBP in the Snf5F/FGFAP-Cre mouse (B). There is also a reduction in the total number of cells in the disrupted corpus callosum of the Snf5F/FGFAP-Cre mouse (C) as well as a decreased expression of the Beta III tubulin stain (D). Scale bars represent 600 m (top) and 200 m (bottom).
Supplemental Figure S2. Snf5/Baf47 expression in Snf5F/F p53L/L GFAP-Cre mutant brain tissue
Higher magnification of H&E (left) and Baf47 (right) at 600x of the lesion in the Snf5F/F p53L/L GFAP-Cre brain (originally illustrated with blue arrows in Fig. 3C (H&E) and Fig. 3D (Baf47) demonstrates a proliferation of highly atypical malignant cells with numerous mitoses and apoptotic bodies.Blue stain indicates loss of Baf47 expression in the subpial area of the molecular layer of the cerebellum (right). Scale bars represent 20 m.
Supplemental Figure S3.NG2 expression in Snf5F/F p53L/L GFAP-Cre mutant brain tissue
(A) Sagittal brain sections of the hippocampus (top) and cerebellum (bottom) of a 3-week old control mouse (left) and a Snf5F/F p53L/L GFAP-Cre mouse (right) with double immunofluorescent labeling of Baf47 (green) and NG2 (red). Scale bars represent 200 m. (B) Sagittal brain sections of the hippocampus (top) and cerebellum (bottom) of an adult control mouse (left) and Snf5F/F p53L/L GFAP-Cre mouse (right) with double immunofluorescent labeling of Baf47 (green) and NG2 (red). Scale bars represent 200m.
Supplemental Figure S4.Olig 2 is significantly reduced in Snf5F/F p53L/L GFAP-Cre adult mutant mice
(A) Sagittal brain sections of the cerebellum of an adult control mouse (left) and a Snf5F/F p53L/L GFAP-Cre mutant mouse (right) with double immunofluorescent labeling of Baf47 (green), Olig 2 (red) and DAPI stain (blue). Scale bars represent 100 m.
(B) Higher magnification of the cerebellum of an adult control mouse (left) and a Snf5F/F p53L/L GFAP-Cre mutant mouse (right) with double immunofluorescent labeling of Baf47 (green), Olig 2 (red) and DAPI stain (blue). Scale bars represent 50 m.
(C) Histogram of Olig 2 counts per square micron in 3 adult control cerebellum (blue) and 3 adult Snf5F/F p53L/L GFAP-Cremutant cerebellum (red). Error bars represent standard deviation and *** indicate p < 0.001. The p value (0.0002788) was calculated using the student T-test in Excel.
Supplemental Figure S5. Gene Set Enrichment Analysis shows close resemblance between human and mouse brain tumors
(A) Two gene sets were generated using the top 100 genes exhibiting the greatest differential expression between human AT/RT and human medulloblastoma RNA as well as that obtained from a comparison of mouse AT/RT and mouse medulloblastoma. Gene set enrichment analysis (GSEA) was carried out using these differential expression gene sets. The results confirmed that genes with higher expression in human medulloblastoma have generally higher expression in the corresponding mouse model, and vice versa. The plots show the overall skewness of differential expression of the group of genes.
(B-D) GSEA was carried out independently, using the differential expression results obtained from the human tumor comparison and the mouse model comparison, against the MSigDB collection of gene sets( The selected gene sets exhibit a close parallel in both the human and mouse data, generally indicating higher expression in AT/RT. The selected gene sets in B and C have generally higher expression in AT/RT in both human and mouse. The selected gene sets in D have generally higher expression in medulloblastoma in both human and mouse.
Supplemental Table S1.Primer sequences of qRT-PCR
This list comprises the primer sequences used in qRT-PCR assays to quantitate gene expression. The table also lists the Assay ID, gene symbol, gene name, context sequence and target exon for each.