Figure 1 | Scientific Reports

Figure 1

From: Phosphorylation of SKAP by GSK3β ensures chromosome segregation by a temporal inhibition of Kif2b activity

Figure 1

Identification of a novel SKAP-Kif2b interaction in mitosis.

(a) SKAP complexes were immunoprecipitated from HeLa cell extract with anti-SKAP antibody or control IgG. The isolated complexes were separated using a gel fraction followed by Western blotting with corresponding antibodies. HC, heavy chain. (b) Co-immunoprecipitation of exogenous SKAP and Kif2b. Extracts from HEK293T cells, transiently co-transfected with FLAG-SKAP and GFP (lane 3) or GFP-Kif2b (lane 4), were incubated with an anti-FLAG mouse antibody agarose beads. The immunoprecipitates were resolved using SDS-PAGE followed by Western blotting analyses. Upper panel, GFP blot; lower panel, FLAG blot. (c) SKAP binds specifically to Kif2b. Purified GST-SKAP proteins were used to isolate GFP-Kif2b or GFP-Kif2c from HEK293T cell lysates. The isolated proteins were fractionated by SDS-PAGE followed by CBB staining (lower) and anti-GFP blotting analysis (upper). (d) Representative immunofluorescence images of mitotic cells expressed GFP-Kif2b were fixed and stained with SKAP (red). Scale bars, 10 μm.

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