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Fig. 5. Developmental expression of Filia. (A) The relative
abundance of Filia and Mater transcripts in mouse oocytes,
eggs and preimplantation embryos. Poly (A)+ RNA was isolated from
oocytes/eggs/embryos, reverse transcribed with oligo (dT) and aliquots were
analyzed by qRT-PCR using synthetic oligonucleotide primers and Taqman probes
specific to each of the two Filia isoforms and Mater. The
amount of 1.2 kb Filia in mid-sized oocytes (50 µm) was set as
1.0. Inset shows primer efficiencies over four orders of magnitude of
substrate, which were used to calculate PCR efficiencies for the two
Filia isoforms and Mater. (B) The persistence of
FILIA and MATER protein during oogenesis and preimplantation embryos.
Immunoblot of lysates isolated from growing oocytes (50 µm, 75 µm),
eggs, two-cell embryos, morulae and blastocysts (E3.5) was incubated with
peptide-purified antibodies that bind to either MATER (upper) or to both
isoforms of FILIA (lower).
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