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Short Report
Trichoplein controls microtubule anchoring at the centrosome by binding to Odf2 and ninein
Miho Ibi, Peng Zou, Akihito Inoko, Takashi Shiromizu, Makoto Matsuyama, Yuko Hayashi, Masato Enomoto, Daisuke Mori, Shinji Hirotsune, Tohru Kiyono, Sachiko Tsukita, Hidemasa Goto, Masaki Inagaki
Journal of Cell Science 2011 124: 857-864; doi: 10.1242/jcs.075705
Miho Ibi
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Peng Zou
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Akihito Inoko
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Takashi Shiromizu
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Makoto Matsuyama
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Yuko Hayashi
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Masato Enomoto
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Daisuke Mori
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Shinji Hirotsune
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Tohru Kiyono
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Sachiko Tsukita
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Hidemasa Goto
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Masaki Inagaki
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  • For correspondence: minagaki@aichi-cc.jp
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Data supplements

  • JCS075705 Supplementary Material

    Files in this Data Supplement:

    • Supplemental Figure S1 -

      Fig. S1. Trichoplein is also localized on desmosomes and keratin IFs. (A,B) Localization of trichoplein (Tricho.; green) and desmoplakin (DP) 1 (red) in mouse small intestine. Nuclei were also stained with DAPI (D; blue). (C) Calcium switch experiments. Cells were treated as described in the legend to Fig. 1D. Treated cells were subjected to immunocytochemistry with anti-Odf2 and anti-desmoplakin (DP) 1/2 at the indicated time after switching. Arrowheads indicate the centrosome position. (D) Trichoplein localization in HeLa cells. HeLa cells were stained with antibodies to pan-keratin or trichoplein. High magnification views are also indicated in insets. Arrowhead indicates a centrosome. Scale bar: 10 µm.

    • Supplemental Figure S2 -

      Fig. S2. Complex formation between His-tagged trichoplein and GFP-tagged Odf2β/ninein in COS7 cells. COS7 cells transfected with vectors carrying indicated proteins were subjected to immunoprecipitation with anti-GFP (A), or affinity chromatography with Ni-NTA-agarose (B).

    • Supplemental Figure S3 -

      Fig. S3. Interaction between trichoplein and Odf2β/ninein. (A) Yeast two-hybrid interaction of trichoplein with keratin-18 (K18), ninein, or Odf2β. (B) GST pull-down assays. 1, MBP + GST-ninein; 2 and 5, MBP-trichoplein + GST; 3, MBP-trichoplein + GST-ninein; 4, MBP + GST-Odf2β; 6, MBP-trichoplein + GST-Odf2β.

    • Supplemental Figure S4 -

      Fig. S4. Effects of trichoplein depletion on cell cycle profile. HeLa cells were treated with control (Cont.) or human trichoplein-specific siRNA (Sq. 4 or 2) for 48 hours. Treated cells were subjected to immunoblotting (left) or FACS analysis (right). As a loading control, cell lysates were also immunoblotted with anti-p38 MAPK (left). For FACS analysis showing the DNA content in each group, about one million treated cells were collected by trypsinization, resuspended in buffer solution (CycleTESTTM PLUS kit; Becton-Dickinson, San Diego, CA) and stored at −80°C. Then, we treated cells according to the manufacturer�s protocol (CycleTESTTM PLUS kit) and analyzed them using a Becton-Dickinson FacsScan and CellQuest software. The percentage of G0−G1, S, or G2−M phase in each group is indicated in the box. We obtained a similar FACS profile with Sq. 4 (data not shown).

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Short Report
Trichoplein controls microtubule anchoring at the centrosome by binding to Odf2 and ninein
Miho Ibi, Peng Zou, Akihito Inoko, Takashi Shiromizu, Makoto Matsuyama, Yuko Hayashi, Masato Enomoto, Daisuke Mori, Shinji Hirotsune, Tohru Kiyono, Sachiko Tsukita, Hidemasa Goto, Masaki Inagaki
Journal of Cell Science 2011 124: 857-864; doi: 10.1242/jcs.075705
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Short Report
Trichoplein controls microtubule anchoring at the centrosome by binding to Odf2 and ninein
Miho Ibi, Peng Zou, Akihito Inoko, Takashi Shiromizu, Makoto Matsuyama, Yuko Hayashi, Masato Enomoto, Daisuke Mori, Shinji Hirotsune, Tohru Kiyono, Sachiko Tsukita, Hidemasa Goto, Masaki Inagaki
Journal of Cell Science 2011 124: 857-864; doi: 10.1242/jcs.075705

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