1. Normal Anaplastic astrocytoma Glioblastoma Figure S1; Soumya et al., 2009 Malignant astrocytoma -3.00.03.0 Figure S1. One-way hierarchical clustering of significance analysis of microarray identified grade specific miRNAs that differentiate malignant astrocytoma from normal brain samples. Normalized, log 2-transformed expression ratios of significance analysis of microarrays identified differentially regulated miRNAs were subjected to one-way hierarchical clustering using TMEV software to see if gene set can cluster samples of different grade separately. A dual color code was used with red and green indicating up- and down-regulation, respectively, in the particular astrocytoma sample compared to normal brain tissue. There were 55 miRNAs found to be up regulated and 29 miRNAs down regulated in malignant astrocytomas in comparison to normal brain samples at a false discovery rate (FDR) of 0.0%.

2. Anaplastic astrocytomaSecondary glioblastoma Primary glioblastoma Figure S2; Soumya et al., 2009 -3.00.03.0 A B Figure S2: Identification of tumor grade specific miRNA. A and B) One-way hierarchical clustering of significance analysis of microarrays identified grade specific miRNAs that differentiate anaplastic astrocytoma and secondary glioblastoma (A) and anaplastic astrocytoma from primary glioblastoma (B). A dual color code was used with red and green indicating up- and down-regulation, respectively. Anaplastic astrocytoma

3. -4 -2 0 2 4 6 8 10 U138U251U343U373 U87 LN18 LN229 Log 2 ratio Normal brainGlioma cell lines miR-21 miR-23a -10 -8 -6 -4 -2 0 2 4 6 U138U251U343U373 U87 LN18 LN229 Normal brain Glioma cell lines SVG -3 -2 0 1 2 3 4 U138U251U343U373 U87 LN18 LN229 Normal brainGlioma cell lines SVG miR-218 miR-219 -2 0 1 2 3 4 U138 U251 U343U373 U87 LN18 LN229 SVG Normal brainGlioma cell lines -2 0 1 2 -3 miR-155 miR-16 U138 U251 U343 U373 U87 LN18 LN229 SVG Glioma cell lines Normal brain miR-25 miR-93a miR-106b Log 2 ratio A B C DE Figure S3; Soumya et al., 2009

4. Figure S3: A: Real time RT-PCR analysis showing the expression of miR-218 and miR-219 in control brain samples (seven) and glioma derived cell lines (eight). B: Real time RT-PCR analysis showing the expression of miR-21 and miR-23a in control brain samples (six) and glioma derived cell lines (seven). C: Real time RT-PCR analysis showing the expression of miR-25, miR-93 and miR-106b in control brain samples (seven) and glioma derived cell lines (eight). D: Real time RT-PCR analysis showing the expression of miR-155 in control brain samples (seven) and glioma derived cell lines (eight). E: Real time RT-PCR analysis showing the expression of miR-16 in control brain samples (seven) and glioma derived cell lines (eight).