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Blood, Vol. 109, Issue 6, 2597-2603, March 15, 2007
Bmi-1 is induced by the Epstein-Barr virus oncogene LMP1 and regulates the expression of viral target genes in Hodgkin lymphoma cells
Blood Dutton et al.
109: 2597
Supplemental materials for: Dutton et al, Vol. 109, Issue 6, 2597-2603
Files in this Data Supplement:
- Table S1. Genes changed by BMI siRNA (XLS, 1.98 MB)
- Table S2. Differentiation related genes (XLS, 177 KB)
- Figure S1. Bmi-1 is not up-regulated by LMP1 expression in Burkitt lymphoma cell lines (JPG, 14.2 KB)
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Immunoblotting analysis shows that induced expression of LMP1 in the EBV-negative BL cell lines BJAB and DG75 did not up-regulate Bmi-1 expression. However, LMP1 was able to up-regulate the expression of TRAF1, an established target of LMP1 in BL cells. These data suggest that the up-regulation of Bmi-1 by LMP1 might be restricted to HL cells.
- Figure S2. Bmi-1 knockdown using an exogenously supplied oligonucleotide siRNA (JPG, 24.3 KB)
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We observed similar effects on gene expression and on cell viability after the knockdown of Bmi-1 using an oligonucleotide siRNA specific for Bmi-1 that targeted a different region of Bmi-1. (A) Immunoblotting for ATM in L428 cells 48 hours after transfection with either the Bmi-1–specific siRNA or a scramble control siRNA compared with cells treated with transfection reagent alone. ATM is up-regulated in cells transfected with Bmi-1 siRNA but not in control cells. Similar effects were seen in the L591 cell line (data not shown). (B) Viability of L428 cells was decreased after transfection of L428 cells with the Bmi-1–specific oligonucleotide siRNA but not of control cells. Similar results were obtained for L591 cells (data not shown). These data confirm the specificity of the effects observed with the separate Bmi-1 knockdown construct reported in Figures 4 and 7.
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