XB-IMG-82319
Xenbase Image ID: 82319
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Figure5. Myo10 Helps Position the Spindle but Functions Antagonistically to F-Actin(A) Side-view immunofluorescent images of spindles in control morpholino (Ctrl MO) and Myo10 MO-injected embryos.(B) 3D reconstructions of single cells from epithelium of Ctrl MO and Myo10 MO embryos.(C) Quantification of spindle position in Ctrl MO, Myo10 MO (both coinjected with GFP as a control), and Myo10 MO rescued with full-length GFP-tagged Myo10 (GFP-HIQT), tailless Myo10 (GFP-HIQCC), or headless Myo10 (GFP-IQT). Spindles are repositioned closer to the apical cell surface in Myo10 MO embryos compared to Ctrl MO, a phenotype rescued by coinjection with full-length or headless Myo10, but not tailless Myo10. To test for significance, unpaired Student's ttests were performed (n= 5 independent experiments for Ctrl MO+ GFP and Myo10 MO+ GFP, from a total of 27 and 32 embryos, respectively; n= 3 independent experiments, from a total of 18 embryos each for Myo10 MO+ GFP-HIQT,+ GFP-HIQCC, and+ GFP-IQT; p< 0.05, p< 0.01, p< 0.01).(D) Quantification of spindle position in Ctrl MO, Myo10 MO (both treated with 0.1% DMSO as a control), and Myo10 embryos treated with Noc or LatB. Treatment with low-dose Noc does not affect Myo10 MO spindle position, but LatB treatment of Myo10 MO embryos causes spindles, on average, to move basally and results in a wider spread of spindle position. To test for significance, unpaired Student's t tests were performed (n= 3 independent experiments, from a total of 20, 16, 18, and 11 embryos for Ctrl MO, Myo10 MO, Myo10 MO+ Noc, and Myo10 MO+ LatB, respectively; p< 0.05, p< 0.01).(E) High-resolution side-view confocal images (stacks of 13z slices for each condition) of spindle microtubules in Ctrl MO, Myo10 MO, and Myo10 MO rescued with GFP-HIQT. In Ctrl MO cells, spindle microtubules have an apical asymmetry, with more astral microtubules on the apical side (arrows). Spindles in Myo10 MO cells lose this asymmetry, and long basal astral microtubules are seen (arrows). The GFP-HIQT rescue restores the apical asymmetry.(F) Filament tracing of the microtubule signal (white trace of red staining) provides an unbiased approach to view the asymmetry of the microtubule network. For each image, the trace represents only microtubules present in the central mitotic cell: any traces originating in neighboring cells were deleted. In particular, a dense network of microtubules is seen on the apical side of Ctrl MO spindles, which is lost in the Myo10 MO and restored in the GFP-HIQT rescue (squarebracket).ns, not significant. Scale bars represent 10m. See also FigureS3. Image published in: Woolner S and Papalopulu N (2012) © 2012 ELL & Excerpta Medica. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license
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