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Additional Supporting Information may be found in the online version of this article.

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DVDY_22417_sm_suppfig-S1.tif19078KSupp. Fig. S1. The expression pattern of Sdf1 and E-cadherin (E-cad) is not changed in the hindgut tissues of the E9.5 CKO embryos. A, B: Whole-mount views of bright-field image. C–C′,′ D–D″: Magnified views of the boxed areas in A and B respectively. C, D: Bright-field confocal images. C′, D′: Fluorescence images of immunostaining for E-cad. C″, D″: Fluorescence images of immunostaining for Sdf1α. A, C–C″: Lhx1 heterozygous (het) embryos. B, D–D″: CKO embryos. Scale bars = 100 μm.
DVDY_22417_sm_suppfig-S2.tif1512KSupp. Fig. S2. PGCs stay away from ectopic Ifitm1-expressing cells in the endoderm. A–C: Lipofection of the vector into the endoderm layer of the E7.5 embryos, and monitoring the expression of the vector by (A) GFP expression. EGFP expression indicates cells expressing the vector in the lipofected embryos. A′, B, C: AP-positive PGCs after 24 hr of in vitro culture. AP-positive PGCs are excluded from the definitive endoderm in D,E CMV-Ifitm1-Ires-EGFP vector transfected embryo (n=2), but not so in (F>) CMV-Ires-EGFP control vector transfected embryo (n=3). Inset in A′ shows a magnified view of the boxed area. Plane of the section in E is shown in A′ (black line). Arrows in B mark the PGCs that remain in the mesoderm of the CMV-Ifitm1-Ires-EGFP vector transfected embryo. Arrowheads in C mark the PGCs that populate the definitive endoderm in the control embryo. en, endoderm; mes, mesoderm. D–F: Electroporation of the vector into the posterior endoderm of E8.25 embryos, and monitored for vector expression by (D, E′, F′) GFP expression and (E, F) localization of AP-positive PGCs after 30 hr of in vitro culture. E, E′: AP-positive PGCs were excluded from the segment of the embryonic gut expressing the CMV-Ifitm1-Ires-EGFP vector, but (F, F′) intermingled with cells expressing CMV-Ires-EGFP control vector in the embryonic gut. E, E′: Magnified views of the boxed area in A. Arrowheads (E–F′) mark the vector-expressing cells. hg, hindgut endoderm.

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