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Fig. 3. eIF4E shifts from a 80 kDa complex in unfertilized eggs to a high-molecular mass complex following fertilization. (A) Eluted fractions from the Sephacryl-200 gel filtration purification of unfertilized (top) or 30 minutes post-fertilization (bottom) extracts were applied to a m7GTP column and bound proteins were subjected to western blotting using anti-human eIF4E and anti-human 4E-BP2 antibodies. Arrows at the top of the figure correspond to the position of molecular mass standards (in kDa) used to calibrate the gel filtration column and the numbers at the bottom correspond to the fraction from the gel filtration purification showed in Fig. 2A. (B) Quantification of the results obtained from unfertilized (open circle) or 30 minutes post-fertilization (filled square) extracts. eIF4E amounts in each fraction were expressed as a percentage of the total amount of eIF4E contained in unfertilized or 30 minutes post-fertilization extracts, respectively. Arrows at the top of the figure correspond to the position of the molecular mass standards (in kDa) used to calibrate the gel filtration column showed in Fig. 2A. These data are representative of more than two independent experiments.





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