Figure 6.

Influence of KLF4 and PGAM1 on Endoderm or Mesoderm Differentiation (HES3)

(A) Normalized gene expression of KLF4 in undifferentiated hESCs (HES3) and sorted CXCR4⁺ DE cells. Values are mean ± SEM, n = 4.

(B and C) Normalized protein expression of KLF4 (B) and quantification of CXCR4⁺ DE cells by flow cytometry (C) after suboptimal DE differentiation and transfection with the indicated miRNA mimics (HES3). Values are mean ± SEM, n = 4. ANOVA plus Bonferroni's post hoc test, ^∗p < 0.05, ^∗∗p < 0.01 compared with hESCs and ^##p < 0.01 between miR-1263 and the NC on the particular day. A representative western blot of KLF4 is depicted in (B).

(D) Normalized gene expression of KLF4 in hESCs and after transfection with the indicated siRNAs plus subsequent suboptimal DE differentiation. Values are mean ± SEM, n = 4–6.

(E and F) Representative flow-cytometric histogram of CXCR4⁺ cells at day 4 (E) and quantification of CXCR4⁺ cells at days 2 and 4 (F) upon siRNA transfection and subsequent suboptimal DE differentiation. Values in (F) are mean ± SEM and one respective NC of each experiment is scaled to 100%, n = 4–6. Student's t test, ^∗p < 0.05.

(G–I) Normalized gene expression of PGAM1 (G), flow-cytometric quantification of PDGFRA⁺ cells (H), and normalized gene expression of PDGFRA and KDR (I) at day 5 of ME differentiation. The cells were reseeded on day 2 and transfected with the indicated siRNAs. Values are mean ± SEM, n = 4–6. Student's t test, ^∗p < 0.05.

See also Figure S6.