Fig. 2. Transfected NTmCENPF redistributes in COS-7 cells expressing GFP-syntaxin 4. (A) COS-7 cells singly transfected with NTmCENPF show a cytoplasmic distribution of the protein (red) with a high perinuclear distribution. (B) Cells singly transfected with GFP-syntaxin 4 (green) show a significantly different distribution at defined foci throughout the cell. (C-E) When cells are cotransfected with syntaxin 4 and NTmCENPF, syntaxin 4 remains at the multiple intracellular foci, and NTmCENPF is redistributed to the same foci occupied by GFP-syntaxin 4, as seen in the merged image (E). DAPI (blue) was used to visualize the nuclei. (F) COS-7 cells were transfected with NTmCENPF-myc and GFP-syntaxin or with GFP-syntaxin 4 alone for a negative control. An immunoprecipitation was conducted with -myc antibody, and blots were probed with -GFP antibody. Input lanes show transfected protein expression in the lysate. GFP-syntaxin 4 was precipitated in the presence of NTmCENPF-myc. The singly transfected control shows there was no spurious binding of GFP-syntaxin 4 to the beads.