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Fig. 6. Examination of mouse ES cells for differentiation ability of
Rex1-/Oct3/4+ cells selected with
thymidine kinase gene/gancyclovir system into primitive endoderm lineage.
(A) Thymidine kinase gene knock-in ES cell lines under selection for
puromycin and gancyclovir (GANC). (Left) Cells selected with puromycin
(selection for Oct3/4+ cells); (right) cells selected with
both puromycin and GANC (selection for Oct3/4+ and
exclusion of Rex1+ cells). (B) Expression of marker
genes was examined by RT-PCR in puromycin and GANC/puromycin double-selected
fractions. (C) Expression of primitive endoderm marker genes in
differentiated Rex1+, Rex1- and `reverted'
Rex1+ cells by withdrawal of LIF. Before starting
induction of differentiation, Rex1+ cells were selected
with blasticidin S whereas Rex1- cells were selected with
GANC and puromycin for 1 week. Reverted Rex1+ cells were
derived from Rex1- cells re-seeded and cultured without
GANC for 3 days, and then cultured with blasticidin S for 4 days. Scale bar:
100 µm.
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