Supporting information for Chestukhin et al. (2003) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0730733100
Fig. 7.
Specificity of separase-specific mAb XJ-13. (A) Endogenous separase was detected by XJ-13 mAb in HeLa cells. After fixation and permeabilization, the cells were stained with XJ-13 mAb (Control) or with XJ-13 pretreated with an excess of the GST-separase fusion used for the antibody production (Blocked). (B) Separase was detected by XJ-13 mAb either in control oligonucleotide-transfected (Control) or in separase-specific ASO 7601 transfected (Anti-Sense) HeLa cells. Centrosomal localization of separase was scored in prometaphase and metaphase cells in reference to the γ-tubulin signal. The percentage of separase-positive (Sep.+) prometa-, and metaphase cells is shown for control and antisense oligonucleotide-transfected cells (Right). One hundred cells were scored for each transfection, and representative cells are shown. DNA was visualized by 4',6-diamidino-2-phenylindole (DAPI) staining. “Merge” shows the overlap of the separase, γ-tubulin, and DAPI staining, where yellow highlights overlap of separase and γ-tubulin signal.