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. 2017 Mar 6;216(3):657–674. doi: 10.1083/jcb.201507113

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© 2017 Saunders et al.

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Figure 3. — LAP1 is required for rearward nuclear movement during centrosome orientation. (A) Representative epifluorescence images of centrosome orientation in NIH3T3 fibroblasts treated with siRNA. A LAP1-depleted cell expressing EGFP-LAP1^WT is shown (arrowhead and inset). (B) Centrosome orientation in the cells described in A. (C) Mean centrosome and nucleus positions in the cells described in B. n ≥ 139. (D) Representative montages of DIC images of nuclear movement during centrosome orientation in NIH3T3 fibroblasts treated with NC or LAP1.1 siRNA. A DIC montage of LAP1-depleted cells expressing EGFP-LAP1^WT (inset) is provided. Nuclei are outlined by dashed white lines. (E) Percentages of the cells described in D with rearward-moving nuclei. (F) Mean velocities of nuclear movement from cells described in D. (G) Representative epifluorescence images of centrosome orientation in LAP1-depleted NIH3T3 fibroblasts expressing the indicated EGFP-LAP1^X construct (arrowheads and insets). Asterisks indicate oriented centrosomes. (H) Centrosome orientation in the cells described in G. (B and H) Dashed yellow lines denote random centrosome orientation. (I) Mean centrosome and nucleus positions in the cells described in G. n ≥ 11. Bars, 10 µm. Error bars show SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.