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Fig. 4. Genetic evidence for the role of laminin 1 in the cell-to-matrix interaction required for epiblast polarization. Upon co-cultivation, GATA4 transformed endoderm-like ES cells rescue the dnFgr mutant (A-D). The GATA6 transferred homozygous Lamc1 mutant, in contrast to the heterozygote (E-H), does not accumulate BM proteins and cannot rescue dnFgfr cells. (A-D) Plastic-embedded sections; (E-H) ß-galactosidase and Neutral Red staining. (E-H) Whole-mount confocal microscopy: green, phalloidin (a marker for F-actin); red, laminin {alpha}1. (A) AB2.2, wild-type EBs; (B) dnFgfr EB expressing ß-galactosidase; (C) GATA4 transformed AB2.2 cells; (D) co-cultivation of dnFgfr (1C6) ES cells with GATA4 transformed AB2.2. (E) GATA6-transformed Lamc1-/- cells. Lamc1-/- GATA6 cells form aggregates with minimal BM protein expression. (F) Co-cultivation of Lamc1-/- GATA6 cells with dnFgr cells does not lead to phenotypic rescue. (G) Lamc1+/- GATA6 cells form cysts containing BM proteins. (H) Lamc1+/- GATA6 cells rescue differentiation of 1C6. Scale bars: 50 µm.





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