Supplementary information
From the following article:
Bone morphogenetic proteins inhibit the tumorigenic potential of human brain tumour-initiating cells
S. G. M. Piccirillo, B. A. Reynolds, N. Zanetti, G. Lamorte, E. Binda, G. Broggi, H. Brem, A. Olivi, F. Dimeco and A. L. Vescovi
Nature 444, 761-765(7 December 2006)
doi:10.1038/nature05349
Supplementary Methods
Detailed description of methods used in this study.
Supplementary Tables
Supplementary Tables 1–9 containing comprehensive data on all cell preparations used.
Supplementary Legends
This file contains legends for Supplementary Tables and Supplementary Figures.
Supplementary Figure 1
A quantitative analysis (Real Time PCR (RT-PCR)) of the expression of the BMPR type 1A, 1B, and 2 genes in the different GBM cell preparations investigated in this study – acutely dissociated or briefly cultured (GBSC) cells.
Supplementary Figure 2
a, Dose response curves showing the ability of BMP4 to inhibit the proliferation of two representative GBM cell preparations in a dose-dependent fashion. b. Cytofluorimetric analysis showing the phosphorylation and nuclear translocation of phospho Smad 1,5,8 as elicited 1.5 hours post BMP4 treatment in cultured GBM cells.
Supplementary Figure 3
Effects of various BMPs (all at 100 ng/ml) on the growth of GBM cells.
Supplementary Figure 4
The figure shows the effect of BMP4 on the expansion rate of briefly cultured GBM cells expanding in the presence of mitogens.
Supplementary Figure 5
The figure shows one example of the dot plot graph concerning the quantitative analysis of the expression of the CD133 antigen.
Supplementary Figure 6
Untreated GBM cells constitutively express neuronal markers such as MAP2.
Supplementary Figure 7
BMP4 inhibits the tumorigenicity of GBM cells.
Supplementary Figure 8
Ex vivo pre-treatment of GBM cells with BMP4 inhibits ventricular invasion following orthotopic injection.
Supplementary Figure 9
Phenotype analysis confirmed that, as shown previously, tumours induced by transplanting briefly cultured GBM cells into the brain of scid/bg animals generated astroglial-like cells.
Supplementary Figure 10
a. Quantitative RT–PCR analysis of the expression of 13 different BMPs genes in briefly cultured GBM cells. b. When GBM cells, cultured with mitogens, were exposed to a neutralizing antibody that specifically blocks BMP4, the cell growth rate increased significantly as compared to control conditions.
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