Xenbase Image ID: 127296

Figure 1. Fuz is essential for distal, but not proximal, axonemal identity. (a, left) A representative control axoneme expressing RFP-CLAMP (top) and pixel intensity along the axoneme (bottom). (right) A collection of individual control axonemal intensity plots. (b, left) A representative axoneme from a Fuz KD cell and its corresponding intensity plot. This image is magnified compared with the image in a to facilitate comparison. (right) A collection of intensity plots from individual Fuz KD axonemes. Arrowheads indicate ectopic CLAMP enrichments. (c) Cilia on a control vertebrate multiciliated cell expressing RFP-CLAMP to label the distal axoneme and GFP-MAP7 to label the proximal axoneme. (d) Cilia on a Fuz KD vertebrate multiciliated cell expressing RFP-CLAMP and GFP-MAP7. Also see Fig. S1 (a and b). The mean percentage of axoneme length ± SD occupied by the CLAMP or MAP7 domains is indicated on the bottom right of c (middle [n = 54] and right [n = 130]) and d (middle [n = 52] and right [n = 130]). See Fig. S1 (d and e) for quantitation; only axonemes whose whole length was obvious were used for this analysis, and the compartment analysis for this dataset is shown in Fig. S1 g. (a–d) Bars, 3 µm. (e) Schematic representations of control and Fuz KD axonemes (Axo) showing mean proximodistal organization (for quantification, see Fig. S1 [d, e, and g]). Note that in Fuz KD axonemes, the MAP7 domain is of approximately the same length but makes up a greater percentage of the overall shorter axoneme. Also, note the reduction in both length and percentage of occupancy of the CLAMP compartment. Lengths are reported as the mean across the population, and the percentage of occupancies is reported as the mean of ratios from individual axonemes and is therefore not directly comparable. See Materials and methods for details. (f) Fuz KD axonemes show reduced enrichment of RFP-CLAMP signal in the distal domain over basal axoneme levels, as compared with controls (Ctl). Enrichment Index = (mean intensity compartment)/(mean intensity of an equivalent length of nonenriched axoneme). Fuz KD enrichment index (mean ± SD = 2.17 ± 0.70; n = 113) versus control enrichment index (4.28 ± 1.68; n = 87; P < 0.0001) is shown. Note that only axonemes with discernable distal RFP-CLAMP compartments were used for this analysis. (g) There is no change in the enrichment of the MAP7 compartment between control and Fuz KD axonemes. Fuz KD enrichment index (mean ± SD = 2.64 ± 0.64; n = 102) versus control enrichment index (2.65 ± 0.80; n = 75; P = 0.6508) is shown. Image published in: Brooks ER and Wallingford JB (2012) © 2012 Brooks and Wallingford. 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-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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