Figure 3

eIF4A regulates cell cycle progression and apoptosis. (a) Gene ontology (GO) analysis predictions of the effects of Silvestrol on cancer-related pathways in MDA-MB-231 cells. (b) Cell cycle progression of MDA-MB-231 cells was monitored after treatment with DMSO (left) or Silvestrol (right) for 24 hours. Cells were pulsed with bromodeoxyuridine (BrDU) for 30 minutes prior to harvest. Upon harvest, cells were fixed, stained with anti-BrDU-fluorescein isothiocyanate (FITC) antibody and propidium iodide (PI) and analyzed by fluorescence-activated cell sorting (FACS) (c) MDA-MB-231 cells were synchronized by serum starvation for 24 hours then released from starvation in the presence of DMSO (top panels) or Silvestrol (bottom panels). The cell cycle was monitored over time by BrDU incorporation and PI staining as in Figure 3a. (d) MDA-MB-231 cells were treated with DMSO or Silvestrol for 0, 2 or 6 hours prior to lysis. Lysates were analyzed by Western blotting for proteins indicated. (e,f) MDA-MB-231 cells were treated with DMSO or Silvestrol from 0, 8, 16 and 24 hours prior to lysis; lysates were analyzed by Western blotting for indicated proteins. (g) Tables showing RPKM values from ribosome profiling data for genes implicated in cell cycle progression (top) and apoptosis (bottom).