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Fig. 3. Morphology of EBs grown with or without Tet in LIF-supplemented medium. After culture in LIF-supplemented medium without Tet for induction of exogenous Sox17, the ESCs were allowed to aggregate and were grown in the same medium for 10 days (Tet –). Control EBs were prepared using the same procedure, except that Tet was constantly supplemented to the medium (Tet +). (A,B) Bright-field images. Bars, 500 µm. (C-F) Toluidine-blue staining. Bars, 50 µm (C,D) and 10 µm (E,F). (G-K) TEM images. Boundaries between the PrE-like surface cells and the subjacent cells are shown as broken lines (G,I). The asterisks show the basement membrane-like structure (H,K). Arrows indicate vacuoles (J). Bars, 10 µm (G,H), 2 µm (I,K) and 1 µm (J). (L,M) PAS staining. The inset shows a magnified image (L). Bars, 100 µm. (N,O) Phase-contrast images of EBs grown in gelatin-coated tissue-culture dishes for 1 day. Bars, 300 µm. Two independent clones were examined with similar results (A-O).
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