Figure 6.

Expression of hsΔ200 GRASP65 arrests cells at the G2/M transition in HeLa cells. A cell line expressing the V5-tagged C-terminus of human GRASP65 (hsΔ200-V5) under an inducible promotor (Switch System, Invitrogen) was arrested in S-phase by double thymidine treatment. The second thymidine incubation was carried out in the absence or presence of mifepristone, which is an inducer of hsΔ200-V5 expression. Thymidine was removed to allow progression into mitosis, in the absence or the presence of the inducer. Cells were fixed 13 h after thymidine washout. Cells were processed for immunofluorescence microscopy using antibodies to V5 to detect hsΔ200-V5–expressing cells and phospho-histone H3 to identify mitotic cells. The quantification of four independent coverslips is shown.