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Fig. 3. DOC1R is regulated by phosphorylation. For A,B, immature oocytes were injected with RNA encoding the MYC-DOC1R protein and further cultured for different length of time. Each sample corresponds to a pool of 25 oocytes from the same injection and to oocytes that expressed the mRNA for at least 6 hours. (A) MYCDOC1R regulation during meiotic maturation. MYC-DOC1R mRNA was injected into wild-type oocytes that were collected at different stages of meiotic maturation: immature (lane 1, GV), at 1 hour (lane 2), 3 hours (lane 3), 6 hours (lane 4) and 14 hours (lane 5, Metaphase II, MII) after GVBD. (B) In vitro dephosphorylation of MYCDOC1R from mouse oocyte extracts. Thirty oocytes were injected with MYC-DOC1R mRNA, cultured 14 hours after GVBD, collected and incubated without (lane 1) or with (lane 2) 400 U of {lambda}-phosphatase (PPase + or -). All samples were analyzed by immunoblotting with the anti-MYC antibody. Experiments have been repeated four times.