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. 2010 Oct 14;285(53):41961–41971. doi: 10.1074/jbc.M110.169607

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Expression levels of Lin28 and its regulatory miRNAs are inversely correlated during ES cell differentiation. A, ES cells were differentiated in vitro by spontaneously self-assembling in suspension culture into three-dimensional cell aggregates (EBs). Immunostaining demonstrated that Lin28 was highly expressed in the undifferentiated ES cells. B, pluripotency and differentiation markers during differentiation and EB formation were monitored by real-time RT-PCR. C, Lin28 mRNA expression during EB formation was analyzed by real-time RT-PCR. D and E, Lin28 protein expression during EB formation was analyzed using Western blots. F, the global miRNA expression profile during EB formation was analyzed using a TaqMan miRNA assay. An unsupervised cluster analysis indicated that all four Lin28-regulatory miRNAs grouped together. G, detailed miRNA expression changes during EB formation identified by the TaqMan miRNA assay. All four Lin28-regulatory miRNAs were markedly up-regulated from day 6 of EB formation. H, real-time RT-PCR validations of the TaqMan miRNA assay in two ES cell lines.