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Fig. 2. Serine 353-phosphorylated CDC25B is located at the centrosome. (A) CDC25B RNA interference. Western blot with anti-CDC25B polyclonal antibody, SE96 and anti-actin of total extracts from HeLa cells transfected or not with CDC25B RNAi or control (cont.) scrambled RNAi. (B,C) HeLa cells were fixed and immunofluorescence staining was performed as described previously (Davezac et al., 2000). Cells were also stained with 4'-6 diamino-2-phenylindole (DAPI). (B) Staining with SE96 antibodies of cells in prometaphase and interphase (top panels), and anaphase and interphase (lower panels). (C) Double immunofluorescent staining with SE96 and a {gamma}-tubulin monoclonal antibody of cells in interphase (top panels) and prometaphase (lower panels); SE96 (red), {gamma}-tubulin (green) and DAPI (blue). (D) HeLa C1 expressing a GFP-centrin fusion protein (Piel et al., 2001) were stained with SE96 antibodies; SE96 (red), GFP-centrin (green) and DAPI (blue).





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