XB-IMG-156494
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Fig. 7.
GRP cilia formation in foxn4 and foxj1 mutant embryos. (A-C) Embryos were injected with Foxj1gRNA1 or Foxn4gRNA1 along with Cas9 protein, followed by RNA encoding mRFP (blue) or Cep152-GFP (green) to label membranes and centrioles, respectively. At stage 17/18, the dorsal tissue was dissected away to expose the GRP, fixed, stained for cilia (red) and imaged by confocal microscopy. Typical confocal images are shown of GRP cells in control (A), Cas9/Foxj1gRNA1-injected (B) or Cas9/Foxn4gRNA1-injected (C) embryos. Anterior is up. (D) Scatter plot showing length of GRP cilia in cells from individual embryos injected as indicated. Mean and s.d. are indicated. (E) Cilia positioning in cells along the anteroposterior axis in the GRP was measured in embryos injected as indicated. Each bar shows data from individual GRPs based on data obtained from 10-30 cells, with the percentage of cells with cilia located in the anterior, middle or posterior third indicated. Most GRP cells have a posteriorly localized cilium in control (69±11%) and Cas9/Foxn4gRNA1-injected (69±9%) embryos, but not in Cas9/Foxj1gRNA1-injected embryos (29±9%; P=9×10−7). Image published in: Campbell EP et al. (2016) Copyright © 2016. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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