Abstract
Although Activin/Nodal signaling regulates pluripotency of human embryonic stem (ES) cells, how this signaling acts in mouse ES cells remains largely unclear. To investigate this, we confirmed that mouse ES cells possess active Smad2-mediated Activin/Nodal signaling and found that Smad2-mediated Activin/Nodal signaling is dispensable for self-renewal maintenance but is required for proper differentiation toward the mesendoderm lineage. To gain insights into the underlying mechanisms, Smad2-associated genes were identified by genome-wide chromatin immunoprecipitation-chip analysis. The results showed that there is a transcriptional correlation between Smad2 binding and Activin/Nodal signaling modulation, and that the development-related genes were enriched among the Smad2-bound targets. We further identified Tapbp as a key player in mesendoderm differentiation of mouse ES cells acting downstream of the Activin/Nodal-Smad2 pathway. Taken together, our findings suggest that Smad2-mediated Activin/Nodal signaling orchestrates mesendoderm lineage commitment of mouse ES cells through direct modulation of corresponding developmental regulator expression.
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Acknowledgements
We thank Gaoyang Zhu for technical assistance. This work was supported by grants from the National Natural Science Foundation of China (30930050, 30921004), the 973 Program (2006CB943401, 2010CB833706) to YGC, and grants from the China National Science Foundation (Grant # 30890033, 30588001 and 30620120433), Chinese Ministry of Science and Technology (Grant # 2006CB910700) to JDH.
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Knockdown of Smad2 does not affect mouse ES cell proliferation. (PDF 46 kb)
Supplementary information, Figure S2
Anti-Smad2 antibody characterization in R1 ES cells. (PDF 24 kb)
Supplementary information, Figure S3
Z score of Parametric Analysis of Gene Set Enrichment (PAGE) analysis. (PDF 37 kb)
Supplementary information, Figure S4
De novo prediction of Smad2-associated DNA motifs and cofactors analysis. (PDF 72 kb)
Supplementary information, Table S1
Smad2 binding sites and corresponding genes (XLS 104 kb)
Supplementary information, Table S2
Differentially expressed genes in response to Activin or SB431542 (shown in four tabs) (XLS 2330 kb)
Supplementary information, Table S3
PCR Primers (XLS 26 kb)
Supplementary information, Data S1
Supplementary Methods (PDF 288 kb)
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Fei, T., Zhu, S., Xia, K. et al. Smad2 mediates Activin/Nodal signaling in mesendoderm differentiation of mouse embryonic stem cells. Cell Res 20, 1306–1318 (2010). https://doi.org/10.1038/cr.2010.158
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DOI: https://doi.org/10.1038/cr.2010.158
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