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. 2013 Jan 14;288(9):6166–6177. doi: 10.1074/jbc.M112.418376

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© 2013 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — Overexpression of Ovol2 represses neural conversion and promotes mesendodermal differentiation of ESCs. A, EBs from control GFP-ESCs or Ovol2-overexpressed ESCs were cultured in unbiased differentiation medium for 8 days. The expression levels of Sox1, Pax6, and Six3 were analyzed by qRT-PCR and normalized to the expression of Gapdh. The values for each marker in the control EBs (Ctrl) were designated as 1, respectively. B, double immunostaining for Sox (red) and Oct4 (green; artificial color) proteins and the percentage of Sox⁺Oct4⁻ cells in day 8 EBs described in A. Scale bar, 100 μm. C, the expression levels of T, Mixl1, Flk1, Gata4, Gata6, and Sox17 in day 4 EBs cultured under the conditions described in A were analyzed by qRT-PCR and normalized to the expression of Gapdh. The values for each marker in control EBs were designated as 1, respectively. Error bars, S.E.