Fig. 6. Aberrant numbers of bbs disturbed the microtubular cytoskeleton. (A) Indirect immunofluorescence of control (a-c), N41 (d) and A10 cells (e-g) using antibodies to -tubulin (a,e'-g'), acetylated (c,d) and polyglutamylated tubulin (b,g). (e,f) Corresponding anti-centrin staining. In control cells, microtubules are focused onto the flagellar apparatus (a), which contains two polyglutamylated bbs (b) and four acetylated microtubular bundles (arrows in c). Arrowheads, axonemes. (d) Methanol-permeabilized N41 cells showing bbs (cell 1 and 2) with an incomplete set of aceylated microtubules (2). Some cells showed only a few acetylated microtubules (cell 3), or no staining (cell 4). (e) Additional bbs were often surrounded by microtubules (arrowhead; compare also g and Fig. 3A,c). (f) Cells without dot-like centrin signals often showed dispersed microtubules. (g) Cell with three bbs (arrowheads) as recognized by GT335 and two microtubular asters (g'). Bars, 5 µm. (B, a) Strain D5 expressing GFP-SFA. (b-e) Strain D5 after induction of centrin-RNAi. Cells were stained for centrin (displayed in white color, not shown in a); GFP-SFA is shown in black. Note the defects in the usually cruciated SFA fibers in cells with bbs of low centrin content (arrows in b) and in cells with too many (c,d) or without (e) bbs. Bar, 5 µm.

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