Figure 1.

Membrane Rigidification Prior to Pluripotency Exit

(A) Flow cytometry analysis of pluripotent cells during endodermal induction. A differentiation marker (CXCR4, marker of definitive endoderm cells) and a pluripotent marker (Tra2-49/6E) were used.

(B) Representative GP images of pluripotent and pre-/post-differentiation cells in the absence/presence of MβCD. Pre-/post-differentiation cells were obtained at days 3 and 5 during endodermal induction, respectively.

(C) Statistical analysis of the mean GP during endodermal induction in the absence (gray) and presence (red) of MβCD and the calculated cholesterol level (blue). Estimated membrane composition is schematically illustrated in the lower panel.

(D) Differentiation (CXCR4) and pluripotency marker (tra2-49/6E) is co-plotted with GP (+MβCD) during induction. Error bars originate from three independent experiments (n = 3).

(E) Microarray analysis of SCD1/hydroxymethylglutaryl-CoA reductase (HMG-CR) expression in iPSCs during endodermal induction, which was reanalyzed data of our previous study (Takebe et al., 2013).

(F) Concentration dependence of CAY10566 (inhibitor of unsaturated lipid synthesis, SCD1) and lovastatin (inhibitor of cholesterol synthesis, HMG-CR) on pluripotency. Error bars originate from the SD of the histograms of Tra2-49/6E expression.

(G) Phosphorylated-Smad2/3 expression of pluripotent cells in the presence and absence of fluidic modulators.

(H) Estimated role of SCD1 and HMG-CoA reductase in the maintenance of pluripotency.

Error bars show the standard deviation.