Fig. 1. Expression of a sense-antisense hybrid construct induced centrin deficiency. (A) Design of the sense-antisense hybrid construct of centrin used to induce RNAi in C. reinhardtii. The positions and restrictions sites of the primers are indicated and +1 indicates ATG of the coding region. (B) Western blot of control (lane C) and the RNAi strains A7, A10 and N41. Membrane strips were stained with amido black or developed with anti-centrin (pCen1). (C) Anti-centrin (mAB6.8) staining of methanol-permeabilized control (a) and the centrin-RNAi strains. In the latter, centrin was concentrated at the bbs, and NBBCs were absent. In A10 (c) and especially N41 (d), numerous cells contained no centrin signal. (e) Overlay image of an isolated nucleus of A7 stained with DAPI (black) and centrin (white). Bar, 10 µm. (D) The average number of flagella () and centrin dots () was plotted against the amount of residual centrin as determined by western blotting. The data show that bb and flagellar development depend on centrin.

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