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Blood, Vol. 113, Issue 16, 3744-3753, April 16, 2009
Specific activation of microRNA106b enables the p73 apoptotic response in chronic lymphocytic leukemia by targeting the ubiquitin ligase Itch for degradation
Blood Sampath et al.
113: 3744
Supplemental materials for: Sampath et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 53.6 KB)
- Table S1. Changes in miRNAome in response to LBH589 in CLL cells (PDF, 94.2 KB)
- Figure S1. Response of p53 protein and p73 mRNA to LBH589 (JPG, 58.2 KB)
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(A) Changes in p21 expression in CLL cells, normal or with del17p, in response to treatment with 10 Gy Ionizing radiation for 2h. (B). Changes in p53 expression in CLL cells, normal or with del17p, in response to treatment with 10 nM LBH589 for 36 h.
- Figure S2. Response of p73 mRNA to LBH589 (JPG, 53.9 KB)
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Expression levels of p73 mRNA and protein in 20 CLL cells exposed to 10nM LBH589 for 36h. An induction of 1.5 fold or greater was used as a cutoff. Samples demonstrating induction of p73 mRNA or protein are indicated in bold.
- Figure S3. Correlation between Mcm7 and miR106b induction (JPG, 42.6 KB)
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Samples were exposed to 10nM LBH589 after which the levels of induction of Mcm7 was compared to that of miR106b.
- Figure S4. E2F1 and myc regulate miR106b expression (JPG, 76 KB)
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(A and B) The Mcm7-miR106b promoter constructs were exposed to LBH589 for 4 h and the luciferase expression was measured. (C) the E2F1 and myc levels in a representative sample in response to 10nM LBH589 for 3 or 6h (D) Positive correlation between myc and E2F1 expression and upregulation of Mcm7-miR106b transcripts in CLL cells after exposure to LBH 589 for 3 or 6 h.
- Figure S5. Expression of miR-130a in primary leukemia samples treated with 10 nM LBH589 for 3–6 h (JPG, 15.4 KB)
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- Figure S6. Response of a representative CLL sample to SAHA and MS275 (JPG, 63.6 KB)
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(A and B) Expression of miR106b and Mcm7 in a representative primary leukemia sample exposed to 3 µM MS275 or 3 µM SAHA for 3 and 6 h. (C) Changes in acetylation of histone H3, changes in Itch, p73, caspase-9 processing (CF=cleaved fragment), GAPDH, and annexin V staining in response to MS275 or SAHA for 36 or 48 h.
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