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Blood, Vol. 107, Issue 3, 1124-1132, February 1, 2006
The ETS factor TEL2 is a hematopoietic oncoprotein
Blood Carella et al.
107: 1124
Supplemental materials for: Carella et al, Vol 107, Issue 3, 1124-1132
Files in this Data Supplement:
- Table S1. TEL-2 alters expression of genes involved in apoptosis, cell cycle and cell growth (PDF, 94 KB) -
Bone marrow from 5 FU-treated mice was either mock-transduced (Un-BM) or transduced with MSCV-IRES-GFP (GFP-BM) or MSCV-TEL2-IRES-GFP (TEL2-BM) retrovirus as described in the “Materials and methods” section. After transduction, cells were allowed to recover in culture for 72 hours and were sorted for GFP expression by FACS. RNA of the GFP+ cells and mock-transduced cells was isolated using Trizol (Invitrogen) following the manufacturer’s recommendations.
RNA quality was confirmed by UV spectrophotometry and by analysis on an Agilent 2100 Bioanalyzer. Ten micrograms of total RNA was processed in the St Jude Hartwell Center Core Facility according to the standard Affymetrix protocol (http://www.affymetrix.com/support/technical/manual/expression_manual.affx) and analyzed on the Affymetrix MOE-430A GeneChip array. Signal values, detection calls, and pairwise expression analyses were performed using the default parameters within the statistical algorithm of Affymetrix GCOS software version 1.2. Signal values were scaled to a 2% trimmed mean target value of 500. Stringent selection criteria were applied to identify differential expression associated with TEL2. The following pairwise comparisons were performed: TEL2-BM vs GFP-BM, TEL2-BM vs Un-BM, and GFP-BM vs Un-BM. Initial selection was based on differential expression between TEL2-BM and GFP-BM. Probe sets with a log2 ratio greater than 1 (>2-fold change) plus a “change call” (P < .006, Wilcoxon signed rank test) were selected for further evaluation. To best identify TEL2-associated changes, the initial selection was filtered to exclude probe sets with a less than 2-fold in TEL2-BM vs Un-BM and those with a greater than 2-fold change in GFP-BM vs Un-BM. Probe-set annotations (March 14, 2005) were obtained from the Affymetrix website (http://www.affymetrix.com/analysis/index.affx). Functional classification of genes was performed using the Affymetrix Gene Ontology browser. Biological processes significantly enriched within the TEL2-associated gene list were identified using the chi-square test (P < .001). The changes in expression between TEL2-transduced and vector-transduced bone marrow are shown, corrected for difference in expression between vector-transduced and untransduced bone marrow.
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