Figure 6.

Cyp26a1 limits RA levels and neuroectoderm differentiation during PS-like differentiation

(A) Sox1-GFP and DR5-RARE-Scarlet reporter expression after PS-like differentiation of non-transgenic wild-type (WT) or Cyp26a1^−/− mESCs. Dotted lines: gates fixed according to the non-transgenic mESCs negative control.

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

(C) Scheme to assess the interplay between Cyp26a1-mediated dampening of RA signaling and TGFβ or Wnt signaling.

(D and E) Sox1-GFP reporter expression in Chrd^−/−Nog^−/− (pink) or Cyp26a1^−/−Chrd^−/−Nog^−/− (purple, orange: AGN added after day 3) cells after PS-like differentiation (D) and quantification (E) (n = 3 independent experiments; ^∗∗∗, p < .001; One-way ANOVA followed by Tukey's post hoc test; data represented as mean ± SD).

(F and G) Sox1-GFP reporter expression in Dkk1^−/− (light green) or Cyp26a1^−/−Dkk1^−/− (dark green, orange: AGN added after day 3) cells after PS-like differentiation (F) and quantification (G) (n = 3 independent experiments; ^∗∗∗, p < .001; one-way ANOVA followed by Tukey's post hoc test; data represented as mean ± SD). See also Figure S6.