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. 2021 Aug 4;7(32):eabf7561. doi: 10.1126/sciadv.abf7561

Fig. 7. Loss of eIF4A3 triggers apoptosis in a dual p53/non-p53–dependent manner.

Fig. 7

(A) Starburst plot comparing expression of DE genes regulated translationally by the 80S monosome or the polysomes following sieIF4A3 treatment of U2OS cells. Common targets with monotonic deregulation are shown in orange (R = 0.79, P < 0.01). (B) Resazurin time course survival assay in U2OS deprived of eIF4A3 using two different shRNAs. (C) Percentage of caspase 3/7–positive apoptotic cells following eIF4A3 knockdown in U2OS cells. (D) Resazurin assay of WT or TP53 knockout HCT116 cells following eIF4A3 knockdown. (E) Resazurin assay in U2OS or ddp53-U2OS cells deprived of eIF4A3 with siRNA. (F) Immunoblotting of cleaved PARP in U2OS or ddp53-U2OS cells following knockdown of eIF4A3 ± siBAX [siRNA targeting BAX (BCL2 associated X, apoptosis regulator)] or/and siPUMA. (G) Comparative analysis of FAS protein levels between U2OS or ddp53-U2OS cells measured by Western blot following sieIF4A3 or siCtr treatment. Neocarzinostatin was used a positive control for induction of apoptosis. The numbers show quantitation of FAS/β-actin signal. (H) Starburst plot comparing DE genes between 80S monosome–based translational and the protein level. Green dots indicate genes affected from the transcriptional throughout the translational and protein level (forward). Magenta dots show genes that are regulated transcriptionally but are subjected to extratranslational regulation control (intensified) (R = 0.83, P < 0.001). (I) Starburst plot comparing DE genes between polysome-based translational and the protein level. The color coding is the same as in (H) (R = 0.76, P < 0.001). All data in (B) to (E) are shown as means ± SD; n = 3 biological replicates; **P < 0.01 and ****P < 0.001).