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Trib2 regulates the pluripotency of embryonic stem cells and enhances reprogramming efficiency.

Experimental & molecular medicine (2017-11-25)
Eun Kyoung Do, Jae Kyung Park, Hyo Cheon Cheon, Yang Woo Kwon, Soon Chul Heo, Eun Jung Choi, Jeong Kon Seo, Il Ho Jang, Sang Chul Lee, Jae Ho Kim
RÉSUMÉ

Embryonic stem (ES) cells are pluripotent cells characterized by self-renewability and differentiation potential. Induced pluripotent stem (iPS) cells are ES cell-equivalent cells derived from somatic cells by the introduction of core reprogramming factors. ES and iPS cells are important sources for understanding basic biology and for generating therapeutic cells for clinical applications. Tribbles homolog 2 (Trib2) functions as a scaffold in signaling pathways. However, the relevance of Trib2 to the pluripotency of ES and iPS cells is unknown. In the present study, we elucidated the importance of Trib2 in maintaining pluripotency in mouse ES cells and in generating iPS cells from somatic cells through the reprogramming process. Trib2 expression decreased as ES cells differentiated, and Trib2 knockdown in ES cells changed their colony morphology while reducing the activity of alkaline phosphatase and the expression of the pluripotency marker genes Oct4, Sox2, Nanog and Klf4. Trib2 directly interacted with Oct4 and elevated Oct4 promoter activity. During the generation of iPS cells, Trib2 knockdown decreased the reprogramming efficiency of mouse embryonic fibroblasts, whereas Trib2 overexpression significantly increased their reprogramming efficiency. In summary, our results suggest that Trib2 is important for maintaining self-renewal in ES cells and for pluripotency induction during the reprogramming process.

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Anticorps monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
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Anticorps anti-glycéraldéhyde-3-phosphate déshydrogénase, clone 6C5, clone 6C5, Chemicon®, from mouse
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Ponceau S, BioReagent, suitable for electrophoresis
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Anticorps monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin, buffered aqueous glycerol solution
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Normal Goat IgG, This Normal Goat IgG is validated for use in ELISA Flow Cytometry Immunoblotting Immunofluorescence Immunohistochemistry Immunoprecipitation for the detection of Goat IgG, Non-immune.