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Fig. 2. Differentiation course of Gsc+ mesendoderm cells analyzed by FACS. (A) Expression of ECD and GFP from day 4 to day 6. The earliest GFP+ cells were detected on day 3.5 and co-expressed ECD (data not shown). On day 4, more than 90% of GFP+ cells are ECD+. Two major populations of GFP+ cells, ECD+ and ECD-, were observed from day 5. (B) Expression pattern of ECD and {alpha}R of GFP+ cells. The two molecules are co-expressed in the day 4 GFP+ cells but segregate into ECD+{alpha}R- and ECD-{alpha}R+ cells on day 6. (C) The FACS pattern of ECD, Vegfr2 and ßR of GFP+ cells on day 5. Vegfr2 and ßR are expressed in the ECD- population exclusively. (D) Cell sorting and re-culture experiments of Gsc+ cells. ECD+ and ECDlow populations in day 4 GFP+ cells were purified and re-cultured for 1 day in SFM containing activin. ECDhigh population could give rise to both ECD+ and ECD- cells.





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