XB-IMG-156488
Xenbase Image ID: 156488
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Fig. 1.
MCC differentiation is disabled in Foxn4 morphants. (A-C) Xenopus embryos were injected at the two-cell stage with a Foxn4 morpholino, followed with RNA encoding mRFP (blue) and Hysl1-GFP (green) to label membranes and centrioles, respectively. At stage 26, embryos were fixed and cilia (red) stained with an acetylated tubulin antibody. Shown are representative confocal images of the skin in a wild-type control (A) and Foxn4 morphant (B) embryo, along with the percentage of MCCs (±s.d.) displaying defective cilia based on scoring eight fields from different embryos. Scale bars: 10 μm. (C) Plot showing the average number of MCCs (based on cilia extension), outer cells (OC) and putative ionocytes (Inc) in a 0.13×0.13 mm field of the skin, based on data from eight or nine stage 26 embryos. Error bars indicate s.d. (D-G) Confocal images of the skin in control (D,F) or Foxn4 morphants (E,G) at stage 26, where basal bodies are marked by expression of Centrin4-GFP (green, D,E) or Chibby-GFP (green, F,G) and cell boundaries with mRFP. (D-G′) Below each image is a z-scan, where the apical surface and basal body positions in wild-type embryos are marked by the upper and lower dashed lines, respectively. (H,I) Confocal images of the skin in Foxn4 morphants, where cell boundaries are labeled with mRFP (blue), basal bodies with Centrin4-GFP (green), and cilia by acetylated tubulin immunostaining (red), fixed at stage 26 (H) and stage 30 (I), along with the percentage of MCCs (±s.d.) with undocked basal bodies or defective cilia extension obtained by scoring at least eight fields from different embryos. In wild-type controls, MCCs with these phenotypes were rarely observed. (J) Scatter plot of basal body number located within 1 μm of the apical surface in wild-type MCCs or those in Foxn4 morphants at the indicated stages. The central bar indicates the mean and error bars indicate s.d. ***P<0.001; n.s., not significant. 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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