Fig 4.

The rate of protein synthesis is increased in m4EHP- and GIGYF2-depleted cells. (A) Immunoblotting of 4EHP and GIGYF2 in HeLa cells transfected with control, m4EHP, or GIGYF2 siRNAs. β-Actin was used as a loading control. The doublet observed on the topmost Western blot corresponds to m4EHP (33). (B) Effects of m4EHP or GIGYF2 silencing on protein synthesis. Protein synthesis was measured by [³⁵S]methionine incorporation normalized to the total amount of protein. The value of the control cells was adjusted to 100%. [³⁵S]methionine labeling was carried out in triplicates and in three independent experiments (n = 3). All values represent means ± standard deviations (SD). (C) Control or siRNA-treated HeLa cells were pulse-labeled with [³⁵S]methionine. Newly synthesized proteins were visualized by SDS-PAGE and autoradiography. Equal amounts of sample were loaded. Numbers to the right indicate molecular masses (kDa).