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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Nat Cell Biol. 2010 Jan 17;12(2):164–169. doi: 10.1038/ncb2016

Figure 2. γ-TuRC is recruited to kinetochores by Nup107-160.

Figure 2

(a) HeLa cells were depleted of GCP2 (second row) or Nup160 (third row) by siRNA. The cells were treated with nocodazole for 16 hrs and processed for indirect immunofluorescence with antibodies against GCP2 (second column, green) and kinetochore marker CREST serum (third column, red) and counterstained with Hoechst 33342 dye to visualize chromosomal DNA (left, blue). Inset shows enlarged, merged image of kinetochores. Top row shows HeLa cells transfected with a control siRNA, treated in an identical manner. Scale bar = 5 µm.

(b) HeLa cells were treated as in (a), stained with antibodies against Nup107 (second row, green) and counterstained with Hoechst 33342 dye to visualize chromosomal DNA (top row, blue).

(c) Whole cell extracts from control HeLa cells (left), cells depleted of Nup160 (center) or cells depleted of GCP2 (right) were analyzed by Western blotting with the indicated antibodies to monitor the overall level of depletion through siRNAs.

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