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Version of Record online: 19 AUG 2011
Copyright © 2011 AlphaMed Press
Volume 29, Issue 9, pages 1362–1370, September 2011
How to Cite
Araki, R., Hoki, Y., Uda, M., Nakamura, M., Jincho, Y., Tamura, C., Sunayama, M., Ando, S., Sugiura, M., Yoshida, M. A., Kasama, Y. and Abe, M. (2011), Crucial Role of C-Myc in the Generation of Induced Pluripotent Stem Cells. STEM CELLS, 29: 1362–1370. doi: 10.1002/stem.685
Author contributions: R.A.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, financial support; Y.H.: collection and assembly of data, and data analysis; M.U., M.N., C.T., M. Sunayama, S.A., M. Sugiura, and M.A.Y.: collection and assembly of data; Y.J.: collection and assembly of data, data analysis and interpretation; Y.K.: data analysis; M.A.: conception and design, financial support, manuscript writing, final approval of manuscript.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSEXPRESS July 5, 2011.
- Issue online: 19 AUG 2011
- Version of Record online: 19 AUG 2011
- Accepted manuscript online: 5 JUL 2011 04:14PM EST
- Manuscript Accepted: 12 JUN 2011
- Manuscript Received: 7 JAN 2011
- Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency
Additional Supporting Information may be found in the online version of this article.
|STEM_685_sm_SuppFigS1.eps||3719K||Supporting Information Figure S1. Teratoma formation|
|STEM_685_sm_SuppFigS2.eps||2850K||Supporting Information Figure S2. Germline transmission/competency of 2A iPSCs. (A) F1 mice derived from 2A-4F-1 iPSCs. (B) F1 mice derived from 2A-4F-33 iPSCs. (C) A 90% chimeric mouse (9 days old) generated using the 2A-3F-EGFPtg-4 clone. 2A-3F-EGFPtg-4 iPSCs harbor the EGFP gene under the control of a ubiquitous promoter. (D) Testes from the chimeric mouse shown in (C). (E) Seminiferous tubule from the testes shown in (D).|
|STEM_685_sm_SuppFigS3.eps||2796K||Supporting Information Figure S3. Examples of various levels of chimerism as defined by coat color|
|STEM_685_sm_SuppFigS4.eps||1588K||Supporting Information Figure S4. Chimeric mouse generation test for iPSCs established using another procedure and another mouse strain. (A) iPSCs generated using a retroviral gene transduction system. (B) iPSCs generated from MEFs derived from Nanog-GFP tg mice. p, germline transmission positive; n, germline transmission negative.|
|STEM_685_sm_SuppFigS5.eps||3498K||Supporting Information Figure S5. Abnormal embryos developed from 3F-iPSCs|
|STEM_685_sm_SuppFigS6A.eps||9380K||Supporting Information Figure S6. Blastocysts at 48 hours post-aggregation. (A) 2A-4F-EGFPtg-1.|
|STEM_685_sm_SuppFigS6B.eps||8984K||Supporting Information Figure 6B. (B) 2A-3F-EGFPtg-4.|
|STEM_685_sm_SuppFigS6C.eps||9545K||Supporting Information Figure 6C. (C) 2A-3F-EGFPtg-15. Merged, bright field and fluorescence images are shown.|
|STEM_685_sm_SuppFIgS6D.eps||1003K||Supporting Information Figure 6D. (D)Reduction rate of GFP signals after aggregation. The results of in vitro blastocyst formation assays of 2A-4F-EGFPtg-1, and 2A-3F-EGFPtg-4, -5, -7, -14 and -20 are shown. Twenty embryos were examined for each clone. The differences in the GFP signals between the 24 h later and at 72 h later timepoints are indicated. Standard errors are shown.|
|STEM_685_sm_SuppFigS7Aai.eps||24442K||Supporting Information Figure S7. (A) Bisulfite sequencing analysis of pluripotent marker genes. Data were analyzed using QUMA software. Open circles indicate unmethylated CpGs, whereas closed circles indicate methylated CpGs.|
|STEM_685_sm_SuppFigS7B.eps||1450K||Supporting Information Figure 7B. (B) Real-time PCR analysis of imprinting genes in iPSCs. The data were normalized using Gapdh gene expression. The relative expression levels compared with each transcript in the R1 cells are presented. Standard deviations (SD) are shown (n=3).|
|STEM_685_sm_SuppFigS8.eps||852K||Supporting Information Figure S8. Chromosome analysis of iPSCs. The frequency of cells with 40 chromosomes is shown. Standard errors (SE) are also shown.|
|STEM_685_sm_SuppFigS9.docx||164K||Supporting Information Figure S9. Differentially expressed genes between 2A-4F-iPSs and 2A-3F-iPS-4 screened via the HiCEP transcriptome analysis method. A total of 20,138 transcripts including unknown mRNAs were analyzed and capillary electropherograms are shown. Red and blue lines indicate the expression patterns for 2A-4F-1 and 2A-4F- 33, and for 2A-3F-1, 2A-3F-3 and 2A-3F-4, respectively. The Y-axis indicates the transcript levels and the X-axis indicates the length of the fragments . Duplicate experiments with each cell line were performed. Information for each peak is provided in the table.|
|STEM_685_sm_SuppMovieS1.avi||19940K||Supporting Information Movie S1. Aggregation of 2A-4F-EGFPtg-1 cells|
|STEM_685_sm_SuppMovieS2.avi||19929K||Supporting Information Movie S2. Aggregation of 2A-3F-EGFPtg-4 cells|
|STEM_685_sm_SuppMovieS3.avi||19934K||Supporting Information Movie S3. Aggregation of 2A-3F-EGFPtg-15 cells|
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