Open Access Highly Accessed Research article

MicroRNAs miR-26a, miR-26b, and miR-29b accelerate osteogenic differentiation of unrestricted somatic stem cells from human cord blood

Hans-Ingo Trompeter1*, Janine Dreesen1, Eugenie Hermann1, Katharina M Iwaniuk1, Markus Hafner2, Neil Renwick2, Thomas Tuschl2 and Peter Wernet1

Author Affiliations

1 University Düsseldorf, Medical Faculty, Institute for Transplantation Diagnostics and Cell Therapeutics (ITZ), Düsseldorf, D-40225, Germany

2 Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, Rockefeller University, New York, NY, 10065, USA

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BMC Genomics 2013, 14:111  doi:10.1186/1471-2164-14-111

Published: 19 February 2013

Additional files

Additional file 1:

Comprehensive miRNA expression data from native (d0) USSC SA5/73 and SA8/25 and osteo-differentiated (day 7) SA5/73 and SA8/25. This Excel file contains three separate sheets, “USSC SA573”, “USSC SA825”, and “Deep Sequencing”. In the first two, raw Ct, U6-RNA normalized dCt, ddCt, and 2-ddCt values (corresponding to fold changes of miRNA expression in day 7 osteo vs. native cells, in grey columns) are given for each miRNA analyzed. The sheet “deep sequencing” indicates results from deep-sequencing derived expression analysis of miR-26a and miR-26b in native USSC SA5/73, SA8/25, SA8/77, and SA4/101 and osteo-differentiated USSC SA8/77 and SA4/101. Total sequence reads for the miRNA indicated as well as frequency of specific sequence reads among all sequence reads are given.

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Additional file 2:

Experimental validation of miRNA target gene predictions. Detailed data of validations of CTNNBIP1 (A), DUSP2 (B), HDAC4-1 (C), HDAC4-O1 and HDAC4-O2 (D), SMAD1 (E), SMAD6 (F), TGFB3 (G), and TOB1 (H) are given. The bar above the graphs depict the respective 3-UTRs, with the blue part representing the analyzed fragment, including putative miRNA binding sites. For further description see Figure 2.

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Additional file 3:

Primers and oligonucleotides. This Excel file summarizes primers and sense- and antisense-oligonucleotides used for 3-UTR cloning and qRT-PCR analysis. Sheet “3UTR cloning”: Primers for generation of PCR products from 3-UTRs, corresponding NCBI accession numbers, and sense- and antisense-oligonucleotides used for experimental target validation of predicted target proteins. Restriction sites used for cloning are given in bold as well as sticky and recessed ends synthesized into oligonucleotides. “Ⓟ” denotes the phosphate group added to the 5-end of oligonucleotides. Mutations in oligonucleotide sequences corresponding to the seed of microRNAs are given in red. Sheet “qRT-PCR”: Forward and reverse primers used for RT-reaction.

Format: XLS Size: 35KB Download file

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