Knocking-down the Sestrin genes significantly compromises the translational repression of the translation machinery upon p53 activation. (A) A table showing fold induction of Sestrin1 and Sestrin2 (SESN1 and SESN2, respectively) in nutlin-3a treated cells for the indicated times compared to control untreated cells. (B) MCF-7 cells were transfected with si-control and si-SESN1 and si-SESN2 siRNA oligos, treated with nutlin-3a for 20 h, and subjected to RNA-Seq and Ribo-Seq analyses. The effect of siRNAs on the RNA level and ribosome occupancy of SESN1 and SESN2 is shown. (C) Comparison between changes in translation efficiency measured for genes encoding the translational machinery (ribosomal proteins and translation initiation/elongation factors) in response to nutlin-3a treatment, in cells knocked-down for Sestrin1 and Sestrin2 (right) or treated with control siRNAs (left). P-values calculated using Wilcoxon test. FPKM, fragments per kilobase of mRNA per million reads; si, small interfering.