Xenbase Image ID: 84618

Fig. 2. Expression pattern of ectopic Erp1. (A) Immature oocytes were left uninjected (None) or injected with 100 pg of Myc3-Erp1C583A mRNA with a full-length 3′UTR (+ Myc3-Erp1 mRNA), cultured overnight, treated with progesterone and collected at the indicated times. Oocyte extracts were treated or not with λ phosphatase and immunoblotted with the indicated antibodies. The asterisk and deg denote background protein and Myc3-Erp1C583A degradation products, respectively. + ℗ , hyperphosphorylated Myc3-Erp1; − ℗, dephosphorylated Myc3-Erp1. exo, Myc3-Erp1C583A; endo, endogenous Erp1. (B) Immature oocytes were injected with Myc3-Erp1C583A mRNA as in panel A, subsequently collected at the indicated times (after the mRNA injection) and subjected to immunoblotting with anti-Myc antibody. Oocytes treated with the protein synthesis inhibitor cycloheximide (CHX; 200 μg/ml) after 5 h of mRNA injection as well as in vitro translated (IVT; unphosphorylated) Myc3-Erp1C583A protein were analyzed similarly. Note that Erp1C583A protein was transiently synthesized within 1 h of mRNA injection, largely degraded within the next 1 h and then persisted stably (see CHX treatment) until at least 12 h of mRNA injection. This result indicates that the ectopic Erp1C583A protein (and its degradation products) detected in immature oocytes and maturing oocytes before GVBD (see panel A) was derived from the one that had been transiently and leakily synthesized immediately after injection of the mRNA. − ℗, (nonphosphorylated) Myc3-Erp1C583A; deg, Myc3-Erp1C583A degradation products. Image published in: Ohe M et al. (2007) Copyright © 2007. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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