Fig. 2.

OGT overexpression alters the abundance of few proteins that are associated with the spindle and midbody. (A) HeLa cells infected with GFP were cultured in SILAC medium containing no heavy isotope; cells infected with OGT were cultured in SILAC medium containing heavy isotope. After purification and mass spectral analysis, the pie chart displays the ratios of (OGT spindle and midbody/GFP spindle and midbody sample) protein abundance for the more than 700 proteins identified in the MS analysis. Eight percent of identified proteins show increased abundance of more than 200% and 1% show a decrease of more than 50% in abundance. (B)The 700 proteins at the spindle and midbody represent numerous different functional classes as categorized by UniProt. (C) Proteins with a 200% increase in abundance upon OGT overexpression fall into multiple functional classes. (D) Molecular motor proteins and adaptor proteins constitute large proportions of the proteins that decrease in abundance by greater than 50%. (E) Schematic of the chromosomal passenger protein complex that associates with OGT and OGA. (F) Immunoblot (IB) analysis of HeLa synchronized by thymidine-nocodazole from cells overexpressing either GFP or OGT shows that overexpression of OGT causes a decrease in the abundance of several proteins in the chromosomal passenger protein complex. (G) Immunoblotting confirms that the abundance of several proteins is unchanged in cells overexpressing OGT.