Fig. 3. Overexpressed GFP-tagged trichoplein is associated with K8/18 filaments in HeLa cells. (A) pEGFP-trichoplein vector was transfected into HeLa cells. The transfected cells were fixed 16 hours later and stained with K18 antibody, tubulin antibody or rhodamine-conjugated phalloidin. The merged images are also shown. (B) HeLa cells expressing GFP-trichoplein were treated with 10 ng/ml demecolcine (Dem.) for 100 minutes, or 5 µM cytochalasin B (Cyt.) for 15 minutes at 37°C. After the treatment, cells were fixed and stained with K18 antibody, tubulin antibody or rhodamine-conjugated phalloidin. (C) In some cells, GFP-trichoplein existed not only around nuclei but also in the cell periphery, where GFP-trichoplein markedly colocalized with K18. Higher magnification images (lower panels) of the areas indicated by arrows and merged images are also shown. Bar, 10 µm.