Fig 5.

Endogenous SPF45 is phosphorylated by MAP kinases in cells. (A) Cell lysates from ovarian benign and cancer cell lines were immunoblotted as indicated. (B) SKOV-3-Myc-SPF45 cells were either grown adherent (Adh) or put in suspension (Sus) culture in the presence or absence of U0126. Myc immunoprecipitates were immunoblotted for phospho-SPF45 and Myc. Cell lysates were immunoblotted for endogenous ERK and p-ERK. (C) A2780 cells were serum deprived overnight and then treated with either serum, DMSO (DM), anisomycin, PMA, H₂O₂, or UV light for 15 min or 60 min. Cell lysates were immunoblotted for endogenous proteins with the indicated antibodies. (D) Serum-starved (SF) A2780 cells were pretreated with DMSO (D), U0126 (UO), SB203580 (SB), or SP600025 (SP) prior to stimulation with either PMA or H₂O₂ for 1 h. Cell lysates were immunoblotted for endogenous proteins as indicated. (E) A2780 cells were put in serum-free medium (SF) overnight and treated with either DMSO (D), U0126 (UO), SB203580 (SB), or SP600025 (SP) for 30 min prior to stimulation with UV light. Cells were harvested 1 h later. Cell lysates were immunoblotted for endogenous proteins as indicated.