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Fig. 1. PGCs of AKT-MER transgenic mice. (A) AKT-MER transgenic mice
(right) and littermate control mice (left) at E11.5. As the Akt-Mer
cDNA is linked to IRES-EGFP, the transgenic mice can be identified by
EGFP fluorescence. (B) Flow cytometry analysis of cells from the gonads
of E11.5 AKT-MER mice. Suspensions were stained for the PGC-specific marker
SSEA-1. EGFP fluorescence was detected in the majority of the transgenic PGCs.
(C) Level of AKT activation in the cultured PGCs. The E11.5 PGCs were
seeded onto SCF-expressing Sl/Sl4-m220 feeder cells and cultured
with or without bFGF and 4OHT for 1 day. The cells were stained with the
SSEA-1 (green) and anti-Ser437-phosphorylated-AKT (pAKT) (red) antibodies. The
fluorescence intensity of phospho-AKT in the individual PGCs was measured
using LSM5 PASCAL confocal microscopy. Relative fluorescence per cell is shown
in the right panel (mean±s.e.m.). The background fluorescence level was
measured in the samples that were not treated with pAKT antibody. Phospho-AKT
signals were stronger in the 4OHT-treated transgenic PGCs than in the
untreated transgenic and the bFGF-treated wild-type controls
(*P<0.01, #P<0.05, Student's
t-test). The signals were stronger in the transgenic PGCs than in the
control PGCs, even in the absence of 4OHT and bFGF
(#P<0.05).
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