Fig. 2. Localization of Ct-eIF4H. (A) Western blot analysis of cytoplasmic and nuclear fractions of C. tentans diploid cells. The cells were biochemically fractionated into a cytoplasmic (C) and a nuclear (N) fraction. The fractions were analyzed by western blotting, using anti-Ct-eIF4H antibodies. The majority (about 95%) of Ct-eIF4H is present in the cytoplasmic fraction, but a small amount (about 5%) is found in the nuclear fraction. The extra band in the cytoplasmic fraction (approximately 3% of the signal in this lane) was specific for the antibody, but we do not know the identity of this band. In lanes 3 and 4, the fractions were probed with an antibody specific for a cytoplasmic protein to check for contamination of cytoplasm in the nuclear extract. (B) C. tentans diploid cells were stained with anti-Ct-eIF4H antibodies and detected by immunofluorescence (left panel). A predominant cytoplasmic staining was seen. Consistently, a weak granular staining was detected in the nucleus. DNA, in the same cell, stained with DAPI (right panel). Bar, 1 µm.

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