Figure 1

Translation is regulated in LPS-activated moDCs. moDCs were stimulated with LPS for the indicated timepoints and after harvesting both the maturation status and protein synthesis were monitored in parallel. (A) The surface accumulation of MHC II, CD86 and MHC I, were measured by flow cytometry in immature (black line), 4 h (blue) or 16 h (green) LPS-stimulated moDCs. (B) The rate of protein synthesis was monitored with puromycin incorporation using immunoblots (top). Cells extracts were separated by denaturing electrophoresis and analyzed by western blot with antibody to the phosphorylated form of eIF2α and the S6 ribosomal protein (bottom). Total eIF2α detection is shown for equal loading control. (C) FACS analysis with antibody to puromycin (SUnSET method) to quantify protein synthesis. Data are representative of at least three independent experiments, each derived from a different DC preparation.