Figure 5

Wnt signaling is involved in RA-promoted neural specification. A-C, Monolayer differentiation of 46C ESCs in RA-treated cultures (N2B27 + 10^-8 M RA) exposed to the GSK-3 inhibitor LiCl (10 mM) or to NaCl (10 mM); A, Cultures of monolayer differentiation, shown in phase contrast or stained for Sox1^GFP; B, Percentage of Sox1^GFP+ cells scored by FACS. #, significant difference between inhibitor-treated cultures and NaCl-treated cultures (p < 0.001); C, Cultures of monolayer differentiation, shown in stained for Oct4 or combination of Oct4 and Hoechst; D-F, Monolayer differentiation of 46C ESCs in RA-treated cultures (N2B27 + 10^-8 M RA) exposed to the GSK-3 inhibitor CHIR99021 (3 μM) or to equivalent amounts of DMSO diluents; D, Cultures of monolayer differentiation, shown in phase contrast or stained for Sox1^GFP; E, Percentage of Sox1^GFP+ cells scored by FACS; F, Cultures of monolayer differentiation, shown in stained for Oct4 or combination of Oct4 and Hoechst; G, Western blot analysis of Oct4 at day 5 under monolayer cultures; H, Q-PCR for Sfrp2 during monolayer differentiation. Scale bars: A, 40 μm; C, D, F, 80 μm.