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. 2013 Sep 1;126(17):3916–3926. doi: 10.1242/jcs.126599

Fig. 1.

Fig. 1.

Phosphorylation of ARPP19 at S67 is required for resumption of meiosis and mediates binding of ARPP19 to PP2A-B55δ. (A) Prophase oocytes were stimulated with progesterone and collected at the indicated times following addition of progesterone. Oocyte lysates were analyzed by western blot for Gwl, phosphorylated MAPK (pMAPK), endogenous S67-phosphorylated ARPP19 (pS67-ARPP19) and total ARPP19. eGVBD: early GVBD, corresponding to the very first pigment rearrangements at the animal pole of the oocyte. GVBD: well-defined white spot at the animal pole. Lanes ‘no GVBD’ and ‘eGVBD’ correspond to oocytes collected at 150 minutes, either without pigment modifications or at eGVBD. (B) Meiotic maturation time-course of oocytes stimulated by progesterone (Pg), and those injected with 155 ng of either WT-GST-ARPP or S67A-GST-ARPP. (C) Prophase-arrested oocytes (Pro) were either injected with WT-GST-ARPP (WT) or S67A-GST-ARPP (S67A) then stimulated or not by progesterone (Pg). Oocytes were collected at GVBD and WT- or S67A-GST-ARPP were pulled down from oocyte lysates. GST pull-down fractions were then immunoblotted for S67-phosphorylated GST-ARPP (pS67-GST-ARPP), PP2A-A, PP2A-C, PP2A-B55δ and GST (GST-ARPP).