Liver-enriched transcription factors regulate MicroRNA-122 that targets CUTL1 during liver development†
Version of Record online: 30 JUN 2010
Copyright © 2010 American Association for the Study of Liver Diseases
Volume 52, Issue 4, pages 1431–1442, October 2010
How to Cite
Xu, H., He, J.-H., Xiao, Z.-D., Zhang, Q.-Q., Chen, Y.-Q., Zhou, H. and Qu, L.-H. (2010), Liver-enriched transcription factors regulate MicroRNA-122 that targets CUTL1 during liver development. Hepatology, 52: 1431–1442. doi: 10.1002/hep.23818
Potential conflict of interest: Nothing to report.
- Issue online: 30 JUN 2010
- Version of Record online: 30 JUN 2010
- Manuscript Accepted: 15 JUN 2010
- Manuscript Received: 7 MAR 2010
- National Natural Science Foundation of China. Grant Numbers: 30830066, 30771151, 30870530
- National Basic Research Program from the Ministry of Science and Technology of China. Grant Number: 2005CB724600
Additional Supporting Information may be found in the online version of this article.
|HEP_23818_sm_Suppfig1.tif||12713K||Supplementary Figure 1. The expression of all known LETFs in human (A) and mouse (B) tissues. We search the six families of LETFs in the UCSC Genome Browser by Gene Sorter, and their expression data was downloaded and gathered.|
|HEP_23818_sm_Suppfig2.tif||2314K||Supplementary Figure 2. BLAT result shows the conservation of primary transcript for miR-122 between various animals. Sequence for the query is the known genomic sequence of woodchuck hcr (primary transcript for miR-122). Sequence information see the reference 22.|
|HEP_23818_sm_Suppfig3.tif||4194K||Supplementary Figure 3. 5'RACE results show the transcription start sites of human (Huh7 cell) and mouse pri-miR-122. A. Agarose gel electrophoresis shows the PCR products of 5'RACE. B. Alignment of genomic sequence shows the detailed transcription start sites of pri-miR-122. The translation start sites of pri-miR-122 are underlined. In woodchuck, it located at S1 site. In human (Huh7 cell), pri-miR-122 could start at two sites (S2 and S5), whereas it could start at four sites in mouse (S1, S2, S3 and S4). The numbers indicate the frequency of each site in the sequenced clones. 8 and 12 clones were sequenced for human and mouse RACE products respectively.|
|HEP_23818_sm_Suppfig4.tif||162K||Supplementary Figure 4. qRT-PCR assays show the effect of the miR-122 mimic (122) on the expression of target genes in HepG2 cells. The level of target genes was normalized to GAPDH and then compared to the samples transfected with control mimic (NC)|
|HEP_23818_sm_Suppfig5.tif||4099K||Supplementary Figure 5. Multiple sequence alignment of the binding site for miR-122 within CUTL1 3'UTR in 43 species. This picture is a result of Blat from the UCSC Genome Browser. The sequence for Blat is the 47-bp fragment of CUTL1 gene that was used for target validation (5'-TGG GTT TTG CAG ACC AGG GTT TGT TTA ATA CAC TCC ATT CTA GGC CA -3', it is also shown in Supplementary Table 3.). The putative binding site for miR-122 seed is shown as green box in the picture.|
|HEP_23818_sm_Suppfig6.tif||3831K||Supplementary Figure 6. A. Flow cytometry analysis shows the effect of the miR-122 mimic on the cell cycle of HepG2 cells. Cells were transfected with 50 nM miR-122 mimic (122) or the control mimic (NC) and were collected, stained and analyzed 48 h after transfection. B. Cell growth assay shows the effect of the miR-122 mimic on the proliferation of HepG2 cells. Cells were transfected with 50 nM of the mimics. 24 h following transfection, cells were trypsinized and seeded into 6-well-plates at 3000 cells per well and maintained for 10 days. Then, the cells were fixed and stained by crystal violet.|
|HEP_23818_sm_Suppfig7.tif||680K||Supplementary Figure 7. Western blot assay shows the expression of SRF in developing mouse livers, HCC cell lines as well as the effect of over-expression of miR-122 on SRF protein expression. All the samples used were the same as before.|
|HEP_23818_sm_Supptable.doc||159K||Supplementary Table 1. Real-time PCR primers for miR-122, U6, LETFs and GAPDH. Some primers are different between mouse (M) and human (H) and the different bases are underlined.|
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