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. 2017 Jul 19;16(15):1440–1452. doi: 10.1080/15384101.2017.1338985

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© 2017 Taylor & Francis

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Figure 5. — The phosphorylation of ARPP19 by Cdk1 is not essential for meiosis resumption. A. Prophase-arrested oocytes were injected or not with either unphosphorylated WT-ARPP19 (ARPP) or S28A-ARPP or Cdk1-thiophosphorylated forms of ARPP: WT-ARPP (pCdk1-ARPP) or S28A-ARPP (pCdk1-S28A-ARPP). Oocytes were then stimulated or not with progesterone (Pg). Meiosis resumption was followed by scoring the % of oocytes at GVBD as a function of time. B. Oocytes from panel (A) were collected at the time of GVBD and Cdk1 activation was analyzed by western blotting Cyclin B2, S287-phosphorylated Cdc25 (pS287-Cdc25), phosphorylated MAPK (pMAPK) and total ARPP19 (GST-ARPP). C. Prophase-arrested oocytes were injected with single S67-phosphorylated ARPP (pS67-ARPP), or single Cdk1-thiophosphorylated forms of either WT-ARPP19 (pCdk1-ARPP) or S28A-ARPP (pCdk1-S28A-ARPP) or with double Cdk1 and Gwl phosphorylated ARPP proteins: WT-ARPP19 (pS67-pCdk1-ARPP) or S28A-ARPP (pS67-pCdk1-S28A-ARPP). As controls, oocytes were stimulated with progesterone (Pg). Meiosis resumption was followed by scoring the % of oocytes at GVBD as a function of time. D. Oocytes from panel (C) were collected at the time of GVBD and Cdk1 activation was analyzed by western blotting Cyclin B2, S287-phosphorylated Cdc25 (pS287-Cdc25), phosphorylated MAPK (pMAPK), Y15-phosphorylated Cdk1 (pY15-Cdk1) and total ARPP19 (GST-ARPP).