XB-IMG-127929
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Fig. 3. Dgp71WD120, Dgrip75175 and wac NBs have severe spindle defects, yet successfully complete bipolar asymmetric divisions.(A–D) WT and mutant 3rd instar larval neuroblasts (NBs) expressing Jupiter-GFP were filmed from before NEB to the onset of cytokinesis. Representative WT (A), Dgp71WD120 (B), Dgrip75175 (C) and wac (D) NBs are shown. Time is shown as min:sec. Time point −00:20 represents one time point before NEB; the asterisk (*) marks the time of the metaphase to anaphase transition. Mutant spindles at the metaphase-anaphase transition consist of only a few k-fibre bundles unlike WT cells, where spindle density is higher (third row). Central spindle formation is strongly impaired in mutant spindles (red arrowheads) compared to WT spindles (white arrowheads). (E) Spindle length was measured over time from NEB to the onset of cytokinesis in WT (n = 11), Dgp71WD120 (n = 9), Dgrip75175 (n = 10), and wac (n = 7) NBs. Data was aligned to Anaphase B onset (time = 0 on the graphs). Mutant cells take significantly longer to undergo division, and have significantly longer spindles than WT NBs at Anaphase-B onset. (F) The density of Jupiter-GFP (as a proxy for MT density) was calculated at spindles in WT (n = 11), Dgp71WD120 (n = 9), Dgrip75175 (n = 8) and wac (n = 5) NBs. Spindle Jupiter-GFP intensity is significantly lower in mutant neuroblasts. Significance testing was conducted using a Student's t-test. Scale bar represents 5 µm. Image published in: Reschen RF et al. (2012) © 2012. 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-NonCommercial-ShareAlike license Larger Image Printer Friendly View |