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. Author manuscript; available in PMC: 2014 Jul 1.
Published in final edited form as: Nat Commun. 2014;5:3318. doi: 10.1038/ncomms4318

Figure 6.

Figure 6

The MPF autoamplification loop is independent of ARPP19 phosphorylation at S109.

(a) S109D-GST-ARPP does not inhibit Cdk1 activation induced by okadaic acid injection (OA) or cytoplasmic transfer (CT). Prophase-arrested oocytes (Pro) were injected with 400 ng of WT- or S109D-GST-ARPP (S109D) then induced to mature either with Pg, by injecting OA or by transferring cytoplasm from MII-arrested oocytes (CT). oocytes treated with Pg, injected with OA or CT were previously incubated with IBMX for one hour. Oocytes were collected at time of GVBD and immunoblotted for Gwl, Cyclin B2 upshift (CycB2), Cdk1 phosphorylation at Y15 (pY15-Cdk1), phosphorylated MAPK (pMAPK) and S67-phosphorylated ARPP (pS67).

(b) Prophase-arrested oocytes (Pro) were injected with 400 ng of S67A-, S109A-, S109D-, S109A/S67A- or S109D/S67A-GST-ARPP then induced to mature by transferring cytoplasm from MII-arrested oocytes (CT). Some oocytes were induced to mature with Pg as a control. Oocytes were collected at GVBD and immunoblotted for Gwl, Cdk1 phosphorylation at Y15 (pY15-Cdk1), phosphorylated MAPK (pMAPK), S67 phosphorylation of GST-ARPP (pS67) and GST (GST-ARPP).