Figure 3.

In vitro reconstitution of neural induction by Wnt and TGFβ antagonists

(A) Experimental strategy to assess the impact of TGFβ or Wnt pathway inhibition on fate induction. SB431542 (SB43) inhibits TGFβ receptors and XAV939 (XAV) is a tankyrase inhibitor.

(B) T-TagBFP and Sox1-GFP reporter expression as measured at day 5 by flow cytometry in non-transgenic mESCs and 2KI mESCs (Control), or after inhibition of the TGFβ (+SB43) or Wnt (+XAV) pathways. Dotted lines: gates fixed according to the non-transgenic mESCs negative control.

(C) Quantification of (B) data (n = 3 independent experiments; ^∗, p < .05; ^∗∗, p < .01; ^∗∗∗, p < .001; two-sided unpaired t test; data represented as mean ± SD).

(D) Experimental strategy to assess the impact of knockouts of antagonists of the TGFβ and Wnt signaling pathways.

(E) T-TagBFP and Sox1-GFP reporter expression as measured by flow cytometry after 5 days of PS-like differentiation of wild-type 2KI (WT) or Chrd^−/−Nog^−/− mESCs. Dotted lines: gates fixed according to the non-transgenic mESCs negative control.

(F) Quantification of the data in (E) (n = 3 independent experiments; ^∗, p < .05; ^∗∗, p < .01; two-sided unpaired t test; data represented as mean ± SD).

(G) T-TagBFP and Sox1-GFP reporter expression as measured by flow cytometry after 5 days of PS-like differentiation of wild-type 2KI (WT) or Dkk1^−/− mESCs. Dotted lines: gates fixed according to the non-transgenic mESCs negative control.

(H) Quantification of (G) data (n = 3 independent experiments; ^∗∗∗, p < .001; two-sided unpaired t test; data represented as mean ± SD).
See also Figure S3.