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. 2011 Oct 14;286(49):42758–42769. doi: 10.1074/jbc.M111.228411

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 7. — CENP-W is stabilized by interaction with B23 (A) B23 knockdown induced destabilization of CENP-W. Two different kinds of B23 siRNA or combined siRNA were transfected into HeLa-FLAG-CENP-W stable cells, and the protein level of endogenous B23 and CENP-W was examined by Western analysis. B, stability of kinetochore components upon B23 depletion. Western analysis was performed to monitor the protein stability of several centromere proteins after B23 siRNA treatment. C, immunofluorescence staining of B23-depleted cells. Cellular distribution of B23 and CENP-W was visualized by immunostaining in B23-depleted HeLa-CENP-W cells. D, CENP-W is stabilized by B23. Increasing amounts of B23 construct encoding either full-length or N-terminal 1–113 amino acids were co-transfected with FLAG-CENP-W in 293T cells, and the protein expression level was examined by Western blot analysis. GFP plasmid was also co-transfected and used as a transfection control.