Fig. 2 | Nature Communications

Fig. 2

From: High-resolution characterization of centriole distal appendage morphology and dynamics by correlative STORM and electron microscopy

Fig. 2

3D STORM reveals the DAP’s unique nine-fold distribution pattern. a Two color SIM images illustrating that the DAPs, as well as the SDA protein Odf2 are organized in toroids of various diameters. b A comparative wide-field and 3D STORM analysis of the SDA component Odf2 and of various DAPs reveals that each protein has a unique pattern of distribution. Each STORM image is accompanied with a black and white image showing the averaged signals from all nine appendages. CCDC41, FBF1, and SCLT1 are organized in discrete signals, while Cep164 N terminus (Nt) occupies loop-like densities that touch laterally. Signals are shown from the proximal end. The Z-depth color coding scheme for STORM images is used throughout the manuscript and is illustrated to the right. c A loop-like organization of Cep164 N-termini in mIMCD3 cells without and with primary cilium. Elongated acetylated tubulin signal was used to detect cilia. (d) Characterization of ANKRD26 localization. Left: SIM reveals that ANKRD26 localizes as a toroid around older mother centriole and colocalizes with Cep164. Right: A comparative wide-field and 3D STORM analysis reveals that ANKRD26 organizes as a toroid consisting of irregularly-shaped lobules. Outer and inner diameters in (b, d) are averages ± s.d. Dashed circles in (b, c, d) delineate approximate position of mother centriole microtubules. e A full-length (FL) and three truncated Cep164 constructs were generated to determine the spatial organization of Cep164 on appendages. f The fragments from (e) were expressed in cells depleted of endogenous Cep164, labeled against HA, and analyzed by STORM. The examples of STORM images are shown. The C-terminal, Δ1200, fragment localizes closest the centriole center, indicating a radial Cep164 organization. g Quantification of HA STORM signals.  A median line and upper and lower quartile is presented in box-and-whisker plots. Scale bars: 1 μm for all wide-field images of centrioles and 500 nm for STORM images. (This Figure is associated with Supplementary Figure 2 and 3)

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