Fig. 4. Analysis of the localization of Ct-eIF4H in Xenopus oocytes. (A) Nuclei (N) and cytoplasm (C) were dissected from injected and uninjected Xenopus oocytes. The presence of Ct-eIF4H was analyzed by western blotting (lanes 1-4). The nuclei from injected oocytes were further dissected and the nucleoplasm (lane 5) and the nuclear membrane (lane 6) were analyzed separately. The position of Ct-eIF4H is indicated. The additional bands represent endogenous Xenopus proteins that were present in both uninjected and injected oocytes. (B) Hormone-induced transcription of the HS VTK gene did not increase the nuclear content of Ct-eIF4H. Oocytes, injected with Ct-eIF4H mRNA and the HS VTK gene construct, were dissected and the content of Ct-eIF4H in nuclei (N) and cytoplasm (C) was analyzed by western blotting in the absence (lanes 3 and 4) or in the presence (lanes 5 and 6) of hormone. Uninjected oocytes were analyzed as a control (lanes 1 and 2). As an internal control for the amount of material loaded in each lane, the endogenous BRG-1 protein on the same filter was detected with a specific antibody. Quantification of the signals showed that the amount of material loaded in lane 5 was approximately 1.3 times the amount loaded in lane 3, which explains the apparent difference in strength of the Ct-eIF4H band in these two lanes.

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