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

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Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 3 — (A) Illustration depicting U2OS carrying the double DOX-inducible system (table S4B). Administration of DOX suppresses the endogenous eIF4A3, while inducing the ectopic expression of WT FLAG-eIF4A3. ev, empty vector. (B) p53 and eIF4A3 protein levels measured by immunoblotting in engineered U2OS following DOX-induced eIF4A3 knockdown using two different shRNAs and concomitant ectopic expression of FLAG-tagged eIF4A3. Arrows depict endogenous and ectopically expressed FLAG-eIF4A3. L, low exposure; H, high exposure. (C) Reactome pathway enrichment analysis of DE genes between siCtr- versus sieIF4A3-treated cells (DESeq2). Z score shows the overall expression trend (up- or down-regulation) of the genes included in a specific gene ontology (GO) term. Magenta points refer to cell cycle terms and green points to splicing. Data were produced with ClueGO in Cytoscape using the default parameters. (D) GO biological process (BP) analysis of genes referring to the term RNA metabolism (C) using ClueGO and Cytoscape. (E) Log₂ fold change (log₂FC) in mRNA levels of genes referring to term rRNA processing (D). (F) Starburst plot comparing expression of DE genes following sieIF4A3 or ActD^L treatment of U2OS cells (R = 0.57, P < 0.001). (G) Reactome pathway enrichment analysis in common up-regulated genes among sieIF4A3 or ActD^L (F, points shown in quadrant I). The inset explains the scaling used. TNF, tumor necrosis factor.