Skip to main content
Article thumbnail
Location of Repository

Online supplemental material JCB

By 

Abstract

Timely anaphase by release of Mps1 from kinetochores • Jelluma et al. S1 Figure S1. Mps1 auto-regulates its turnover at kinetochores. (A) Western blot showing faster mobility of inactivated Mps1 compared with active Mps1. (B) LAP-Mps1-WT localization at kinetochores of UTRM10-WT cells treated as indicated, detected by immunofluorescence using anti-GFP antibody to detect the LAP-tag. Graph represents quantitation of fluorescence intensities; error bars represent SEM (per condition n = 5 cells, fluorescence intensities of 22 separate kinetochores per cell were determined). Western blot shows total expression levels of uninhibited and inhibited LAP-Mps1-WT. Note that UTRM10-WT cells express near-endogenous levels of LAP-Mps1-WT (Jelluma et al., 2008a), and levels are not changed upon inhibition, as judged by the levels of the loading control �-tubulin. (C) Mps1 localization in UTRM-LAP-Mps1 M602G cells treated as indicated. Graph shows quantitation of fluorescence intensities at kinetochores; error bars represent SEM (per condition n = 8 cells, fluorescence intensities of 22 separate kinetochores per cell were determined). (D) Phospho-Aurora B intensity levels in UTRM10-WT cells treated as indicated. Graph shows quantitation of fluorescence intensities; error bars represent SD (total cell fluorescence intensities were determined; n = 5 cells [DMSO and ZM447439] or n = 10 cells [Mps1-IN-1]). (E) LAP-Mps1-WT localization at kinetochores of UTRM10-WT cells treated as indicated. (F) Live fluorescence images of U2OS cells transfected with the indicated constructs and treated as indicated. Peroxisomal localization is identifiable by the punctate pattern of fluorescence in the cytosol (Gould et al., 1989). (G) Comparison of FRAP of LAP-Mps1-WT on kinetochores in PtK2 cells treated with nocodazole, MG132, and Mps1-IN-1, and of LAP-Mps1 M602G on kinetochores in UTRM-LAP-Mps1 M602G cells treated with nocodazole, MG132, and 23-dMB-PP1 as measured and shown in Fig. 1 E. (H) Immunolocalization of LAP-Mps1-WT in UTRM10 cells treated as indicated

Year: 2013
OAI identifier: oai:CiteSeerX.psu:10.1.1.351.9978
Provided by: CiteSeerX
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://citeseerx.ist.psu.edu/v... (external link)
  • ftp://ftp.ncbi.nlm.nih.gov/pub... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.