|
Fig. 8
Positioning of MTOCs is affected in MZlurap1 mutants and caJNK-overexpressing embryos. a–l Immunofluorescence confocal microscopic images and statistical analysis on fixed embryos at 90% epiboly. The centriole, nucleus, and cell membrane are labelled by centrin4-GFP, DAPI, and mRFP, respectively. The position of the centriole is determined by measurement of the angle formed by the line connecting the centre of the nucleus and the centriole, relative to the anteroposterior axis passing through the nucleus. The statistical data (Student’s t-test) are presented as radar graphs, and the number on each ring represents the percentage of centrioles. The total number of cells (n) examined from three different batches of embryos is indicated on the bottom right of each graph. a–c Location of centrioles (stars) in neuroectoderm cells of indicated embryos. The images are positioned with anterior (A) on the top, and posterior (P) towards the bottom. d–f Statistical analyses (Student’s t-test) show the positions of centrioles in neuroectoderm cells. The centrioles in WT embryos display biased location at the posterior position of the nucleus. They shift to more lateral region of the nucleus in MZlurap1 and caJNK-overexpressing embryos, with some degrees of left (L) and right (R) symmetry. g–i Location of centrioles (stars) in notochord cells of indicated embryos. j–l Statistical analyses (Student’s t-test) show the preferential location of centrioles at the posterior and lateral positions of the nucleus in notochord cells of WT embryos, and randomised location of centrioles in MZlurap1 and caJNK-overexpressing embryos. m–o Representative images from live time-lapse imaging shows the movement trajectories of centrioles in notochord cells of indicated embryos at 90% epiboly. For convenience, both the nucleus and the cell membrane were all labelled in red, by Histone2B-RFP and mRFP, respectively. Scale bar: a–c, g–i, m–o 20 µm