Extended Data Figure 9: Response of SOX17-knockout comp-hPSC to SOX17.

a, Overexpression of dex-inducible SOX17 (iS) in SOX17-knockout comp-hPSCs. b, Gene expression (RT–qPCR) on day 4 of FACS-sorted NANOS3–mCherry (NC)⁺alkaline phosphatase (AP)⁺ hPGCs by RT–qPCR. c, FACS analysis of day 2 embryoids induced from wild-type comp-hPSCs, SOX17-knockout comp-hPSCs and SOX17-knockout comp-hPSCs rescued with SOX17GR transgene (iS) (percentage of CXCR4⁺AP⁺ cells). d, FACS pattern of day 2 embyoid induced from NANOS3–tdTomato reporter comp-hPSCs showing AP⁺CXCR4⁺ cells expressing NANOS3–tdTomato. e, Represents SOX17-inducible system (iSdd). Expression of SOX17 fused with destabilized domain (DD) can be induced by doxycycline (dox); addition of Shield1 (S1) can stabilize SOX17–DD protein. f, Western blots showing SOX17 expression level in day 5 embryoids from SOX17-knockout + inducible SOX17–DD (iSdd) comp-hPSCs. Embryoids were induced with cytokines. To induce SOX17, different concentration of dox and S1 were added. As controls, NANOS3–mCherry⁺AP⁺ hPGCs and NANOS3–mCherry⁻AP⁻ cells from wild-type comp-hPSC-derived embryoids induced with cytokines were used. Histone H3 (H3) was used for internal control. g, Immunostaining of day 4 embryoids from SOX17-knockout + iSdd comp-hPSCs. Embryoids from SOX17-knockout and wild-type comp-hPSC-induced with cytokines were used as controls. Scale bar, 50 μm. h, Quantification of immunostaining data in Extended Data Fig. 9g. The numbers of OCT4⁺BLIMP1⁺ hPGCs, FOXA2⁺ endodermal cells and OCT4⁺BLIMP1⁺ hPGCs expressing FOXA2 were counted from 3 different embryoids. The proportions of the 3 populations are shown. i, Expression of SOX17 and BLIMP1 (RT–qPCR) in day 4 embyoids in response to different SOX17 dosage.

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