XB-IMG-155168
Xenbase Image ID: 155168
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Figure 3. Cp110 localizes to cilia-forming basal bodies in MCCs.(A–D) Cp110 localizes to cilia-forming basal bodies in Xenopus epidermal (A, C) and human airway epithelial cell (HAEC) (B, D) MCCs. (A) gfp-cp110 (green) was expressed at levels permitting ciliogenesis, together with centrin4-cfp (basal bodies, blue). Immunofluorescent staining (Ac.-α-tub., red) was used to visualize cilia. (B) Immunofluorescent staining for endogenous Cp110 (red), cilia (Ac.-α-tub.; blue) and Actin (green) in MCCs (n donors = 1, n MCCs = 4). Yellow arrows in A’ and B’ indicate the base of cilia. (C) Apical view (top) of individual MCC co-injected with gfp-cp110 (green), centrin4-cfp (blue) and clamp-rfp (red) to visualize Cp110, basal bodies and rootlets, respectively. Localization of basal bodies to the apical membrane is shown in lateral projection (bottom). n embryos/MCCs: (4/18). (C') High-magnification analysis of GFP-Cp110 (green, indicated by yellow arrows and green circle) binding to an individual basal body from the MCC shown in (C) (basal body and rootlet are indicated). Inset shows rootlet domain (dashed box) with increased brightness. (D) Lateral projection of MCC stained for endogenous Cp110 (green) and cilia (Ac.-α-tub.; red). n donors = 1, n MCCs = 12 (same samples as in Figure 3—figure supplement 1C) (E–F) Endogenous Cp110 (green) and Centrin 1 (red) staining shows Cp110 adjacent to MCC basal bodies by confocal microscopy (E) and 3D-SIM imaging (F). n donors = 1, n MCCs = 3 each for confocal and 3D-SIM.DOI:
http://dx.doi.org/10.7554/eLife.17557.010Figure 3—figure supplement 1. Cp110 localizes to cilia-forming basal bodies in MCCs.(A–B) GFP-Cp110 localization to basal bodies in Xenopus MCCs. (A) GFP-Cp110 (green) localizes to basal bodies (Centrin4-CFP, blue) prior to apical docking, during the stages of apical basal body transport. Actin staining shown in red. n = 2 embryos, 18 MCCs. (B) GFP-Cp110 (green) shows asymmetric localization to basal bodies/rootlets (Clamp-RFP, red) along the anterior-posterior axis. n = 3 embryos, 30 MCCs. (C) Immunofluorescent staining for Cp110 (green) and cilia (Acetylated-α-tubulin, red) shows Cp110 localization at the level of basal bodies and at the lateral membrane (white arrows) in human HAEC MCCs. Three levels along apical-basal axis are shown (top, apical ciliary tuft level; middle, apical MCC membrane level; bottom, cytoplasmic level). n = 1 donor, 12 MCCs. (D–E) Mouse trachea staining for Cp110 (green), cilia (Acetylated-α-tubulin, red) and nuclei (DAPI, blue). (n = 4). (D) Magnified view of MCCs. (E) Greater area view of mouse trachea with multiple MCCs. (E’) Negative control immunofluorescent staining as described in (E), but without the use of primary anti-Cp110 antibody. (n = 1).DOI:
http://dx.doi.org/10.7554/eLife.17557.011 Image published in: Walentek P et al. (2016) © 2016, Walentek et al. 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 license Larger Image Printer Friendly View |