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. 2011 Jan 3;286(9):7609–7618. doi: 10.1074/jbc.M110.166603

FIGURE 7.

FIGURE 7.

DUPD1 physically associates with ERK1/2 and p38 MAPK. GG-CL cells were transfected with PRL-RS and treated with PRL (1 μg/ml) for 30 min. A, total protein extracts were subjected to IP with T-ERK1/2 or rabbit IgG, and immunocomplexes were resolved on SDS-PAGE, transferred onto PVDF membrane, and immunoblotted against DUPD1 or T-ERK1/2s. Lanes 1 and 2, IP with IgG (0 and 30 min PRL-treated samples); lanes 3 and 4, IP with T-ERK1/2 (0 and 30 min PRL-treated samples); lanes 5 and 6, pre-IP samples (total protein extracts); lanes 7 and 8, flow through after IP (unbound supernatants). B, cells were IP with DUPD1 antibody or goat IgG, and immunocomplexes were resolved and immunoblotted as described for A. C, cells were immunoprecipitated with T-ERK1/2 antibody or goat IgG, and immunocomplexes were resolved and immunoblotted against CDC25A and T-ERK1/2. D, GG-CL cells were transfected with PRL-RS and treated with PRL for 30 min. Co-localization of T-ERK1/2 (green) and DUPD1 (red) was examined by immunocytochemistry using specific antibodies. Blue, DAPI. E, paraffin-embedded ovary sections from PRLR−/−RS mice (2.5 days pregnant) were analyzed for co-localization of active ERK1/2 (green) and DUPD1 (red).