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. 2013 Sep 26;155(1):135–147. doi: 10.1016/j.cell.2013.08.031

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© 2013 ELL & Excerpta Medica.

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Figure S1 — Generating FUCCI-hESCs for Studying Cell-Cycle-Dependent Events in Live Pluripotent Stem Cells, Related to Figure 2

(A) Representative colony of Fucci-hESCs. Fucci-hESCs fixed with 4% PFA and visualized by fluorescence microscopy. Scale bar, 100 μm.

(B–E) hESCs differentiate nonsynchronously and nonhomogenously. Live unsorted Fucci-hESCs were differentiated into (B) endoderm and (C) neuroectoderm and analyzed for germ layer and pluripotency marker expression at different time points flow cytometry. Gates depict negative FL5- and positive FL5+ populations. (D-E) Early G1 phase directs endoderm whereas late G1 promotes neuroectoderm differentiation. Live Fucci-hESCs sorted into early G1 phase, late G1, G1/S transition or S/G2/M phase cells were differentiated into (D) endoderm or (E) neuroectoderm and analyzed for germ layer and pluripotency marker expression at different time points by flow cytometry.

(F) Overview of the initiation of differentiation in hESCs. Early G1 phase directs cells into endoderm and mesoderm whereas neuroectoderm is blocked. Late G1 phase directs cells into neuroectoderm whereas endoderm and mesoderm is blocked.