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Figure 5


Fig. 5. Reconstitution of complexes of syndapin II with dynamin II at cellular membranes in vivo. (A-L) Mitochondrially targeted full-length syndapin II-l, detected by anti-Flag immunostaining (A,D,G,J), recruits different GFP-tagged splice variants of dynamin II (B,E,H,K) when cotransfected in COS-7 cells, as also demonstrated by merging the fluorescence channels (C,F,I,L). (A-F) Recruitment of dynamin IIaa-GFP (B,E) by mito-syndapin II-l that, in A, was additionally co-localized with MitoTracker (in blue; merge in A thus appears magenta). (G-I) In vivo reconstitution of protein complexes composed of mito-syndapin II-l and dynamin IIab-GFP. (J-L) Mito-syndapin II-l efficiently recruits dynamin IIbb-GFP. (M-O) By contrast, in cells cotransfected with mito-syndapin II-l P480L (M), no such recruitment was observable, but dynamin IIaa-GFP (N) shows a rather diffuse localization. O represents the corresponding merged image. Insets are twofold enlargements of boxed areas. Dyn, dynamin; mito-Sdp, mito-syndapin. Bars, 15 µm.





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