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Blood, Vol. 113, Issue 16, 3754-3764, April 16, 2009
Differentiation stage–specific expression of microRNAs in B lymphocytes and diffuse large B-cell lymphomas
Blood Malumbres et al.
113: 3754
Supplemental materials for: Malumbres et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 74.5 KB)
- Table S1. Primers used for 3′-UTR amplification by PCR (PDF, 33.8 KB) -
Nucleotides added to the primers to allow for endonuclease digestion are in bold.
- Table S2. Primers used for site directed mutagenesis (PDF, 59.1 KB) -
Nucleotides that change from original sequence are underlined. In all cases nucleotides were mutated to their complementary ones.
- Table S3. Nucleotides and accessibility of putative binding sites (PDF, 82.5 KB)
- Table S4. Probe IDs (XLS, 27 KB)
- Figure S1. Correlation between microarray data and TaqMan microRNA assays (JPG, 62.7 KB)
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Log2 ratios (lymphoid cell RNA sample/ reference RNA) of array data are plotted in comparison with log2 of 2exp-ΔCt values obtained from Real Time-PCR TaqMan® MicroRNA Assays (Applied Biosystems, Foster City, CA, USA). RNU6B was used as housekeeping gene to calculate ΔCt for the TaqMan microRNA assays.
- Figure S2. Additional paralog families of miRNA clusters overexpresed in centroblasts (JPG, 41 KB)
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Paralog clusters miR-15b/miR-16-2 and miR-16-1/miR-15a (A) and clusters from the miR-181 family (B). Log ratio values are plotted. Only miRNAs that are the main processed product of the corresponding pre-miRNA and without missing values in the microarrays were included. Dash lines are used to represent miRNAs not included in the 39 miRNAs cell of origin classifier.
- Figure S3. Paralog families of miRNA clusters overexpresed in memory cells (JPG, 62.2 KB)
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Paralog clusters miR-23b/miR-24-1 and miR-24-2/miR-23a (A) and paralog clusters miR-29a/miR-29b-1 and miR-29c/miR-29b-2 (B). Log ratio values are plotted. Only miRNAs that are the main processed product of the corresponding pre-miRNA and without missing values in the microarrays were included. Dash lines are used to represent miRNAs not included in the 39 miRNAs cell of origin classifier.
- Figure S4. Regulation of the 3′ UTR of BCL2 transcript by hsa-miR-15a and hsa-miR-15b (JPG, 13.8 KB)
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HeLa cells were seeded the day before transfection, 40,000 cells per well in 24 well plates. Transfection was performed with Lipofectamine 2000 (Invitrogen Gibco BRL, Grand Island, NY) following manufacturer instructions and using 120 pmols of the corresponding pre-miR (Ambion, Austin, TX), 2.5 ng of pRL-TK plasmid and 0.25 µg of the BCL2 3′ UTR pGL3 control construct per well. Dual luciferase ratios are normalized as percentage of the transfection with the non-targeting control pre-miR negative control #1 (Ambion, Austin, TX) (white bar). The mean of 3 independent experiments performed by triplicate is shown. Error bars are the Standard Error of the Mean.
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