Figure 3.

Functional Analysis of Endoderm-Specific miRNAs (HES3)

(A) Schematic overview of the transfection of miRNA mimics/inhibitors.

(B) Time courses of the derivation of CXCR4⁺ cells during suboptimal DE differentiation after transfection with mimics or inhibitors. Values are mean ± SEM, n = 5–6. ANOVA plus Bonferroni's post hoc test, ^#p < 0.05 or ^##p < 0.05 (miR-489-3p), ^∗∗p < 0.01 (miR-1263), ^$$p < 0.05 (both) compared with the negative control (NC) on day 2 or day 4, respectively.

(C) Representative flow-cytometric histograms of CXCR4 staining at days 2 and 4.

(D and E) Normalized expression of marker genes for pluripotency (POU5f1, SOX2) (D) and primitive streak/DE (GSC, FOXA2) (E) after 4 days of differentiation post transfection. Values are mean ± SEM, n = 5–6. ANOVA plus Bonferroni's post hoc test, ^∗p < 0.05 compared with the respective NC.

(F) Schematic overview of the transfection of miRNA mimics/inhibitors under hESC cultivation condition.

(G and H) Normalized expression of marker genes for primitive streak/DE (SOX17, FOXA2, GSC) (G) and pluripotency (POU5f1, SOX2, NANOG) (H) 2 or 4 days after transfection and cultivation in hESC medium. Values are mean ± SEM, n = 4.

See also Figures S4A–S4D.