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Fig. 2. Full length and truncated ODF2 proteins form fibrillar structures in vivo partially associated with microtubules. Blue, DAPI counterstain; green, ODF2-GFP fluorescence; red, immunolocalization of 
-tubulin. C,F,J,M,P,S are merged images of ODF2-GFP, -tubulin and DAPI stainings. (A-G) Localisation of ODF2NC in cos-7 cells. ODF2NC fused to GFP was expressed in cos-7 cells. The protein is present in the nucleus and in the cytoplasm and shows a fibrillar structure (A,G). In G the fluorescence of the nucleus is too bright to discern the centrosome. Detection of microtubules with antibodies against -tubulin (B,C,E,F) revealed partial colocalisation to the microtubule network. Association with microtubules is also demonstrated by colocalisation of ODF2 to the mitotic spindle (D-F, arrows). Arrows in A and G indicate fibrous structures; arrowheads indicate MTOC. Scale bars: 10 µm. (H-S) Localisation of truncated ODF2-GFP fusion constructs transfected in cos-7 cells. Odf2NC1-GFP (H-J), Odf2NC2-GFP (K-M) and Odf2N2C-GFP (N-S) were transfected into cos-7 cells and the microtubule network highlighted by anti--tubulin staining. ODF2NC1 and ODF2NC2 showed cytoplasmic localisation whereas ODF2N2C was preferentially located inside the nucleus. All constructs are found to be concentrated in the centrosome (arrowheads) and have a fibrillar appearance (arrows). Colocalisation to the mitotic spindle is demonstrated for ODF2N2C (Q-S). Scale bars: 10 µm (H,K), 20 µm (N). (T-V) Cytoplasmic organisation of ODF2 is mostly MT independent. Odf2NC-GFP transfected cos-7 cells were treated with Nocodazol (2 µg/ml for 1 hour) for disruption of microtubules. Microtubules were detected by anti--tubulin staining (U,V). Whereas Nocodazol treatment disrupted MT organisation (U), the cytoplasmic ODF2 organisation is not affected (T).
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