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. 2021 Feb 22;2021:6674283. doi: 10.1155/2021/6674283

Figure 1.

Figure 1

Constitutive expression of Lin28a/b in mouse ESC induced Yap1 expression and reduced lineage-specific gene expression. (a) Phase-contrast microscopy and AP staining of Ctrl and Lin28a/b constitutively expressed (Lin28a/b OE) mouse ESCs grown under 2i + LIF medium. Scale bar, 200 μm. (b) Western blot analyses of total proteins from Ctrl and Lin28a/b OE mouse ESCs using the indicated antibodies. Oct4, Sox2, and Nanog are pluripotent stem cell markers. (c) Quantitative real-time PCR to examine the mRNA level of lineage-specific gene expression in Ctrl and Lin28a/b OE mouse ESCs, trophectoderm gene Cdx2, ectoderm gene Nestin, mesoderm gene T, and endoderm gene Gata6. Actin was analyzed as an internal control. The data are shown as the mean ± S.D (n = 3). Statistically significant differences were indicated (∗, P < 0.05 and ∗∗, P < 0.01). (d) Western blot analyses of total proteins from Ctrl and Lin28a/b OE mouse ESCs using the indicated antibodies. (e) Quantitative real-time PCR to examine the mRNA level of Yap1 and its downstream target gene Ctgf expression in Ctrl and Lin28a/b OE mouse ESCs. The data are shown as the mean ± S.D (n = 3).