Figure 3.

Inhibition of NODAL signaling counteracted the expression of NODAL and FOXH1, and the colocalization of FOXH1 and p-SMAD2/3

(A) Sample images and quantification among multiple iPSC cell lines and clones for immunostaining of organoids at day 12. The immunofluorescence imaging showing the spatial expression of FOXH1, p-SMAD2/3, and NODAL in CO treated by olanzapine-treated (Olan), olanzapine+SB431542 dual-treated (Olan+sb), control, and SB431542 single-treated (Control+sb) groups. Scale bar, 20μm. Values represent mean ± SEM (11–30 total rosette structures from at least three organoids).

(B–E) The results of the intensity of p-SMAD2/3 (B), FOXH1(C) the Pearson’s of p-SMAD2/3 and FOXH1 (D) and NODAL (E) between four groups. ^∗∗∗∗p < 0.0001, ^∗∗∗p < 0.001, ^∗∗p < 0.01, ^∗p < 0.05, ns: p > 0.05. ANCOVA adjusting for three cell lines. Post-hoc analyses were conducted using Least Significant Difference (LSD) adjustment method. Data are represented as mean ± SD. (Detailed statistical data can be found in Table S11).