The induction of EMT by depletion of eIF3e negatively regulates the translation of PRP4K in MCF10A cells. A ) Quantitative PCR of PRP4K gene ( PRPF4B ) mRNA expression in control (shCtrl) and eIF3e-depleted (shEIF3E-1 and shEIF3-2) MCF10A cells treated with 10 μg/mL Actinomycin D for the indicated time periods, with data normalized to at least two reference genes as indicated. N = 3, error bars= SEM. Significance was determined by a one-way ANOVA. B ) Western blot analysis of whole cell lysates of MCF10A shCtrl and shEIF3E cells treated with 50μg/mL cycloheximide for the indicated time periods. C ) Puromycin incorporation assay of MCF10A shCtrl and shEIF3E cells. Cells were grown for 24 h without puromycin selection and then pulsed with puromycin prior to Western blot analysis of total cell lysates (left) or following immunoprecipitation (IP) of PRP4K and detection of puromycylation of proteins and PRP4K (right).