XB-IMG-174765
Xenbase Image ID: 174765
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Fig. 3. The MgcRacGAP SxIP motif is necessary for successful cytokinesis and proper astral microtubule structure. (A) Still images from time-lapse movies
of X. laevis embryos at NF stage 10-11. Embryos were co-injected with Mgc MO and either MgcWT or MgcSxNN along with a probe for DNA (mChe–H2B) and
a probe for F-actin (mChe–UtrCH). Red asterisks indicate multinucleate cells. (B) Still images from time-lapse movies of NF stage 13-14 X. laevis embryos injected as
described in A. Red asterisks indicate multinucleate cells. (C) Quantification of the percent of multinucleate cells in MO+MgcWT or MO+MgcSxNN embryos at
NF stage 10-11 or NF stage 13-14. Stage 10-11 – MO+MgcWT=230 cells, 13 embryos, 5 experiments; MO+MgcSxNN=203 cells, 12 embryos, 5 experiments; stage
13-14 – MO+MgcWT=109 cells, 3 embryos, 1 experiment; MO+MgcSxNN=60 cells, 4 embryos, 1 experiment. (D) Still images from time-lapse movies of NF stage
10-11 X. laevis embryos injected as described in A showing symmetric (MO+MgcWT) and asymmetric (MO+MgcSxNN) divisions. Quantification of the percent of
asymmetric cell divisions in MO+MgcWT or MO+MgcSxNN embryos is indicated below each panel. (E) Images of fixed NF stage 10-11 X. laevis embryos that had
been co-injected with a lineage tracer (mChe–membrane, not shown) along with MO alone, MO+MgcWT, or MO+MgcSxNN, then fixed and stained with anti-α-tubulin
antibodies to reveal MTs, and DAPI to label the DNA. (F) Schematic of the astral MT intensity quantification strategy. Intensity measurements were taken from the
boxed regions and averaged for each cell. (G) Quantification of astral MT intensity, normalized to the cytosol background. Note that in order to observe astral MTs, the
spindle MTs sometimes had to be saturated, but the MTs used for astral MT quantification were not saturated. Control=7 cells, 6 embryos; MO=7 cells, 7 embryos;
MO+MgcWT=7 cells, 6 embryos; MO+MgcSxNN=8 cells, 6 embryos. Mean±s.e.m. (unpaired Student’s t-test). Scale bars: 25 µm (A,B); 10 µm (D,E). All n values
represent cumulative values from three independent experiments, unless otherwise indicated. Maximum projection images are shown and were used for
quantification. Image published in: Breznau EB et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, The Company of Biologists Ltd. Larger Image Printer Friendly View |