Figure 2: Expression of markers for the inner cell mass (ICM) and trophectoderm (TE), and for Yap/Tead4 and Notch1/Rbpj in control and Sbno1^Δ/Δ embryos.

(a–l) Immunohistochemistry of ICM (Oct3/4, Nanog) and TE markers (Cdx2, Eomes; Eomesodermin, Krt8; Keratin 8) in E2.5/E3.5 control (a,b,d,e,g,h,j,k) and E3.5 Sbno1^Δ/Δ embryos (c,f,i,l). In 16-cell Sbno1^Δ/Δ embryos at E3.5, Oct3/4 and Nanog were expressed at similar levels to those in the 16-cell stage control embryos (a–f). On the other hand, expression of Cdx2, Eomes and Krt8 was not evident in Sbno1^Δ/Δ embryos (a–c,g–l). (m) RT-PCR analysis of E3.5 control and Sbno1^Δ/Δ embryos. Although Oct3/4, Sox2 and Gata6 were expressed at normal levels, expression of Nanog, Cdx2, Eomes and Krt8 was greatly decreased in Sbno1^Δ/Δ embryos. Hprt was used as an internal control. (n–v) Immunohistochemistry of Yap, Cdx2, Tead4 and Rbpj in E2.5/E3.5 control (n,o,q,r,t,u) and E3.5 Sbno1^Δ/Δ embryos (p,s,v). Nuclear localization of Yap in the Cdx2-expressing TE cells was observed in the control embryos (n,o), In Sbno1^Δ/Δ embryos, expression of Cdx2 was very faint but Yap is clearly localized in nuclei (p). Expression of Tead4 and Rbpj in Sbno1^Δ/Δ embryos was as the same as that in the control (q–s,t-v). Semiquantitative RT-PCR analysis showed expression of Yap, Tead4, Notch1 and Rbpj in control and Sbno1^Δ/Δ embryos at E3.5 (w). Uncropped image of gels are shown in Supplementary Fig. 10.