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Fig. 2. The ninein N-terminus/C-terminus chimerical fusion is concentrated on the mother centriole. (A) L929 cells were transiently transfected with a GFP-tagged construct encoding a ninein version lacking the coiled-coil domain (Nter/Cter-Nin, green). 2 hours after transfection, cells were fixed and centrioles labelled with either GT335 or anti-
-tubulin antibody (red). Representative images show colocalization of centriole markers with the ninein fusion at one centriole. Bars, 10 µm (4 x magnification in insets). (B) L929 cells expressing GFP/Nter/Cter-ninein for 2 hours were fixed and labelled with an antibody to C-Nap1 (a), which decorates the proximal ends of both centrioles (cartoon), or to ninein (b), which essentially decorates the distal end of the mother centriole (cartoon). The GFP signals did not colocalize with C-Nap1 labelling but colocalized with ninein to the mother centriole, showing that GFP/Nter/Cter-ninein is associated with the distal end of the mother centriole or of both centrioles, depending on the cell (C) (same magnification as insets in A). (C) Quantification of Nter/Cter-ninein localization in living cells at 2 hours and 6 hours after transfection. Most cells presented localization on one spot representing the mother centriole or on two spots representing both centrioles.
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