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. 2019 Apr 8;8:e42930. doi: 10.7554/eLife.42930

Figure 2. Notch regulates CDX2 in the early morula.

(A–B) Normalised expression of Cdx2, Gata3, Oct4 and Nanog in pools of 25 embryos treated with RO4929097 to inhibit Notch (left) or with Verteporfin to inhibit YAP/TEAD interaction (right), from the 2 cell to morula stage (A; Notch inhibition, n = 6); YAP/TEAD inhibition, n = 6–9) or from morula to blastocyst stage (B; Notch inhibition, n = 4–6; YAP/TEAD inhibition, n = 6–11). n represents number of unique pools of 25 embryos. Pools of embryos treated with DMSO were used as controls. * p-value<0.05 by Student’s t test. (C) Maximal projections of confocal images after immunostaining of CDX2 in wildtype and Rbpj-/- early morulae. (D) Violin plots of CDX2 intensity levels and (E) quantification of number of CDX2 positive cells in wildtype and Rbpj-/- early morulae (n = 11 embryos). (F) Maximal projections of confocal images after immunostaining of CDX2 in wildtype and Rbpj-/- late morulae. (G) Violin plots of CDX2 intensity levels and (H) quantification of number of CDX2 positive cells in wildtype and Rbpj-/- late morulae (wildtype, n = 4 embryos; Rbpj-/-, n = 5 embryos). Nuclei were stained with DAPI. Scale bar, 20 μm. Data are means ± s.d. ***p<0.001 by Student’s t test (D) or by Fisher’s exact test (E).

Figure 2.

Figure 2—figure supplement 1. CBF1-VENUS levels decrease upon Notch inhibition.

Figure 2—figure supplement 1.

Maximal projections of confocal images of embryos from the CBF1-VENUS mouse line after treatment with RO4929097 to inhibit Notch or with DMSO as control. Treatments were performed in embryos from 2 cell to morula stage (A, n = 6) or from morula to blastocyst stage (B, n = 3). Nuclei were stained with DAPI. Scale bars, 20 μm.
Figure 2—figure supplement 2. Downregulation of CDX2 in Notch1-/- early morulae.

Figure 2—figure supplement 2.

(A) Maximal projections of confocal images after immunostaining of CDX2 in wildtype and Notch1-/- early morulae. (B) Violin plots of CDX2 intensity levels and (C) quantification of the number of CDX2 positive cells in wildtype (n = 11) and Notch1-/- (n = 11) early morulae. (D) Maximal projections of confocal images after immunostaining of CDX2 in wildtype and Notch1-/- late morulae. (E) Violin plots of CDX2 intensity levels and (F) quantification of the number of CDX2 positive cells in wildtype (n = 5) and Notch1-/- (n = 3) late morulae. Nuclei were stained with DAPI. Scale bars, 20 μm. *p<0.05 by Student’s t test (B); ***p<0.001 by Fisher’s exact test (C).