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. 2010 Nov 18;20(4):621–633. doi: 10.1089/scd.2010.0209

FIG. 3.

FIG. 3.

Oct4 negatively regulates Wnt/β-catenin signaling in P19 EC cells. (A) The relative expression levels of primitive streak-specific genes in Oct4-knockdown cells in the presence or absence of β-catenin, as examined by qRT-PCR. One day after transfection, cells are cultured with puromycin for 1 additional day, followed by RNA extraction. (B) β-catenin-dependent activation of Wnt/β-catenin signaling reporter in response to Oct4 knockdown. (C) Enhancement of Wnt/β-catenin signaling by Oct4 knockdown. The basal level of Wnt/β-catenin signaling is elevated by a constitutively active mutant form of Wnt coreceptor Lrp5ΔN. (D) Inhibition of Wnt/β-catenin signaling in a dose-dependent manner by overexpression of Oct4. (E) Overexpression of Oct4 does not impair the activity of FOPFLASH reporter. (F) Inhibition of Wnt/β-catenin signaling by overexpressing Oct4 in HEK293 cells. Wnt/β-catenin signaling is activated by the transfection of plasmid encoding β-catenin. In (B–F), cells are lysed for luciferase assay 1 day after plasmid transfection. (G) Upregulation of various mesoderm genes in 3-day aggregates is diminished by Oct4 overexpression.