Figure 8.
Gemin5 F381A overexpression reverts the effect on the polysome/80S ratio. (A) Polysome profiles (10–50% sucrose gradient) from cytoplasmic lysates of HEK293 cells expressing Xpress-Gemin5 (purple line), Xpress-Gemin5-F381A (pale blue line) and HEK293 control cells (blue line). (Top) Equal amounts of total protein were separated in SDS-PAGE to determine the expression of Xpress-G5 and Xpress-G5-F381A by immunoblotting using anti-Xpress antibody. Tubulin was used as loading control. (B) Representation of the mean polysome/80S ratio obtained from two independent polysome profiles determined in cells transfected with Xpress-G5 or Xpress-G5-F381A compared to control cells. (C) Effect of Xpress-G5 or Xpress-G5-F381A overexpression on global protein synthesis. The intensity of [35S]-methionine labeled proteins during a 3 h pulse was compared to the control cells; values represent the mean ± SD obtained in at least three independent assays. Asterisks denote P-values (*P < 0.1; **P < 0.05, ***P < 0.01).