Figure 8.
Figure 8. Syntelic attachments are stabilized by NOD chimeras that possess either plus end–directed motility or tip-tracking activity. (A) Schematic diagrams of full-length NOD-mCherry and EB1- and kinesin-1-NOD-mCherry. (B) Maximum intensity projection of a GFP–α-tubulin (green)– and EB1-NOD-mCherry (red)–expressing S2 cell showing enrichment of EB1-NOD on chromosomes and the presence of misaligned chromosomes. (C) Maximum intensity projection of a cell expressing Ndc80-GFP (green) and EB1-NOD-mCherry (red) with syntelic (S) and bioriented (B) attachments. (D) Kymograph of a rapid EB1-NOD–mediated chromatin stretch event. (E) Plot of percentage of syntelic attachments versus mCherry fluorescence for EB1-NOD cells. EB1-NOD-mCherry overexpression stabilizes syntelic attachments at a significantly lower frequency than wild-type NOD-mCherry (NOD, n = 57 cells; EB1-NOD, n = 71 cells). (F) Maximum intensity projection of a GFP–α-tubulin (green)– and kinesin-1-NOD-mCherry (red)–expressing cell showing aberrant spindle morphology. (G) Maximum intensity projection of an Ndc80-GFP (green)– and kinesin-1-NOD-mCherry (red)–expressing S2 cell with a mixture of syntelic (S) and bioriented (B) attachments. (H) Kymograph of a persistent kinesin-1-NOD–mediated chromatin stretch event. (I) Plot of percentage of syntelic attachments versus mCherry fluorescence for kinesin-1-NOD cells. Kinesin-1-NOD-mCherry overexpression induces a dose-dependent increase in the percentage of syntelic attachments that rises more slowly and plateaus at a lower percentage of syntelics than the corresponding increase seen for wild-type NOD-mCherry (NOD, n = 57 cells; kinesin-1-NOD, n = 72 cells). (J) The percentage of syntelic attachments in high-expressing cells (defined as >5.0e6 A.U) for NOD-, kinesin-1-NOD-, and EB1-NOD-mCherry–expressing cells. Two-tailed p-values are <0.0005. Error bars are the SEM. Curves were fit with a hyperbolic function. R values are 0.66 (EB1-NOD), 0.7 (kinesin-1-NOD), and 0.86 (NOD). Bars: (B and F) 10 µm; (C and G) 5 µm; (D and H) 1 µm (horizontal) and 10 s (vertical). See Video 10.

Syntelic attachments are stabilized by NOD chimeras that possess either plus end–directed motility or tip-tracking activity. (A) Schematic diagrams of full-length NOD-mCherry and EB1- and kinesin-1-NOD-mCherry. (B) Maximum intensity projection of a GFP–α-tubulin (green)– and EB1-NOD-mCherry (red)–expressing S2 cell showing enrichment of EB1-NOD on chromosomes and the presence of misaligned chromosomes. (C) Maximum intensity projection of a cell expressing Ndc80-GFP (green) and EB1-NOD-mCherry (red) with syntelic (S) and bioriented (B) attachments. (D) Kymograph of a rapid EB1-NOD–mediated chromatin stretch event. (E) Plot of percentage of syntelic attachments versus mCherry fluorescence for EB1-NOD cells. EB1-NOD-mCherry overexpression stabilizes syntelic attachments at a significantly lower frequency than wild-type NOD-mCherry (NOD, n = 57 cells; EB1-NOD, n = 71 cells). (F) Maximum intensity projection of a GFP–α-tubulin (green)– and kinesin-1-NOD-mCherry (red)–expressing cell showing aberrant spindle morphology. (G) Maximum intensity projection of an Ndc80-GFP (green)– and kinesin-1-NOD-mCherry (red)–expressing S2 cell with a mixture of syntelic (S) and bioriented (B) attachments. (H) Kymograph of a persistent kinesin-1-NOD–mediated chromatin stretch event. (I) Plot of percentage of syntelic attachments versus mCherry fluorescence for kinesin-1-NOD cells. Kinesin-1-NOD-mCherry overexpression induces a dose-dependent increase in the percentage of syntelic attachments that rises more slowly and plateaus at a lower percentage of syntelics than the corresponding increase seen for wild-type NOD-mCherry (NOD, n = 57 cells; kinesin-1-NOD, n = 72 cells). (J) The percentage of syntelic attachments in high-expressing cells (defined as >5.0e6 A.U) for NOD-, kinesin-1-NOD-, and EB1-NOD-mCherry–expressing cells. Two-tailed p-values are <0.0005. Error bars are the SEM. Curves were fit with a hyperbolic function. R values are 0.66 (EB1-NOD), 0.7 (kinesin-1-NOD), and 0.86 (NOD). Bars: (B and F) 10 µm; (C and G) 5 µm; (D and H) 1 µm (horizontal) and 10 s (vertical). See Video 10.

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