Research
Myc and Miz-1 have coordinate genomic functions including targeting Hox genes in human embryonic stem cells
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* Corresponding author: Paul S Knoepfler knoepfler@ucdavis.edu
1 Department of Cell Biology and Human Anatomy, University of California Davis School of Medicine, Sacramento, CA, USA
2 Genome Center, University of California Davis School of Medicine, Sacramento, CA, USA
3 Institute of Pediatric Regenerative Medicine, Shriners Hospital For Children Northern California, Sacramento, CA, USA
4 Department of Molecular and Cell Biology, University of California Davis School of Medicine, Sacramento, CA, USA
Epigenetics & Chromatin 2011, 4:20 doi:10.1186/1756-8935-4-20
Published: 4 November 2011Additional files
Additional file 1:
Tables S1 to S5. Table S1. Miz-1 direct targets. Table S2. Myc direct targets. Table S3. Miz-1 and Myc targets. Table S4. Myc knockdown gene expression changes. Table S5. Miz-1 knockdown gene expression changes.
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Additional file 2:
Figure S1. Confirmation of Myc knockdown in human embryonic stem (ES) cells by western blot analysis.
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Additional file 3:
Figure S2. Differentiation of human embryonic stem (ES) cells into embryoid bodies (EBs) leads to a drastic reduction of levels of Myc bound to Hox genes, and a significant upregulation of differentiation-associated genes. (A) Chromatin immunoprecipitation (ChIP) analysis of N-Myc and Miz-1 binding in human ES cells and EBs. (B, C) Real-time quantitative real-time (qRT)-PCR PCR of a series of differentiation-associated genes.
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Additional file 4:
Figure S3. Confirmation of Miz-1 knockdown (KD) in human embryonic stem (ES) cells. (A) Western blot analysis of Miz-1 protein levels. (B) Phase contrast images of human ES cells transduced with either scrambled small hairpin (sh)RNA control or shRNA specific to Miz-1. Images were taken at 4 × magnification.
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Additional file 5:
Figure S4. Myc associates with Miz-1, HDACs, and DNMT3A in vivo in human embryonic stem (ES) cells. (A) Coimmunoprecipitation with a c-myc antibody or a non-specific IgG. Western blot analysis was performed using antibodies specific to Miz-1, Dnmt3a, HDAC1, HDAC2, and HDAC3. TRIM28 was used as a negative control for the interaction with Myc. (B) Chromatin immunoprecipitation (ChIP) on Myc and Miz-1 cobound gene targets.
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