Fig. 3

Localization of GFP-OFD1. a, b Immunofluorescence. Projection of confocal section through the dorsal (a) or the ventral (b) surface of transformants expressing GFP-OFD1 stained by 1D5. a An interphase cell. Right insets the GFP-OFD1 signal (green) overlaps the 1D5 labeling (red) on all basal bodies. Bottom inset an optical transverse section (top outside, bottom inside the cell) performed across the cell surface shows the GFP signal at the distal part of all basal bodies. b A dividing cell. Red inset an optical section through the cell surface at the level of duplicating basal bodies. In addition to the distal GFP labeling (green) at the distal part of the parental basal bodies (red), the GFP signals, not labeled by 1D5, localize at the site where new basal bodies are expected to assemble. Green inset an optical section at the level of the “invariant field.” A dual GFP labeling is detected near the posterior basal body of the doublet when the new basal body assembles: one corresponding to the nascent basal body and the other one corresponding to the anchoring site of the disassembled one. c Immunoelectron microscopy. Localization of the GFP tag after pre-embedding (upper panel) and post-embedding (lower panel) immunolabeling. On longitudinal sections, the labeling is located just above the terminal plate near the intermediate plate (arrows). The gold particles are always localized to the outer side of the axoneme at the level of the microtubular doublets