title: Mechanism of SB431542 in inhibiting mouse embryonic stem cell differentiation.
abstract: SB431542 (SB) is an established small molecular inhibitor that specifically binds to the ATP binding domains of the activin receptor-like kinase receptors, ALK5, ALK4 and ALK7, and thus specifically inhibits Smad2/3 activation and blocks TGF-β signal transduction. SB maintains the undifferentiated state of mouse embryonic stem cells. However, the way of SB in maintaining the undifferentiated state of mouse embryonic stem cells remains unclear. Considering that SB could not maintain embryonic stem cells pluripotency when leukemia inhibitory factor was withdrawn, we sought to identify the mechanism of SB on pluripotent maintenance. Transcripts regulated by SB, including message RNAs and small non-coding RNAs were examined through microarray and deep-sequence experiments. After examination, Western blot analysis, and quantitative real-time PCR verification, we found that SB regulated the transcript expressions related to self-renewal and differentiation. SB mainly functioned by inhibiting differentiation. The key pluripotent factors expression were not significantly affected by SB, and intrinsic differentiation-related transcripts including fibroblast growth factor family members, were significantly down-regulated by SB. Moreover, SB could partially inhibit the retinoic acid response to neuronal differentiation of mouse embryonic stem cells.
authors.affiliation: College of Life Sciences, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Du
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China.
authors.forename: Yongyan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: College of Life Sciences, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Ai
authors.affiliation: College of Life Sciences, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Shi
authors.affiliation: College of Life Sciences, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China.
authors.forename: Linlin
authors.initials: L
authors.lastname: Chen
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China; Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A&F University, 3 Taicheng Road, Yangling 712100, Shaanxi, China. Electronic address: gzk@nwsuaf.edu.cn.
authors.forename: Zekun
authors.initials: Z
authors.lastname: Guo
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004149:Dioxoles / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism; D010766:Phosphorylation; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D053605:Leukemia Inhibitory Factor; D035683:MicroRNAs; D051899:Smad2 Protein; D051900:Smad3 Protein; D005346:Fibroblast Growth Factors; D004121:Dimethyl Sulfoxide
keywords: Differentiation; SB431542; Smad2/3; Undifferentiated state