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. 2021 Jun 8;6(11):e144920. doi: 10.1172/jci.insight.144920

Figure 1. Optimal protein synthesis is necessary for efficient urothelial cell transformation in vivo, and eIF4E phosphorylation selectively increases in the context of bladder cancer formation.

Figure 1

(A) Puromycin incorporation in WT and rpL24+/– urothelium. Representative IF images show less protein synthesis in rpL24+/– mice compared with WT counterparts. Quantification of > 5000 cells/genotype (WT [n = 3], rpL24+/– [n = 2], P < 0.0001, t test). (B) Kaplan-Meier survival analysis of WT (n = 17) and rpL24+/– (n = 13) mice treated with 0.075% BBN ad libitum (P = 0.02, log-rank test). (C) Puromycin incorporation in normal and tumor organoids developed from WT and WT + BBN–treated mice. Representative puromycin Western blot. Quantification of n = 3 biological replicates (P = 0.02, t test). (D) Candidate gene analysis of translation regulators by Western blot using normal and BBN tumor organoids (n = 3 biological replicates). The same tubulin blot is used in the PI3K-AKT-mTOR pathway and integrated stress response figures. The same tubulin blot is used in the translation initiation and translation elongation figures. (E) eIF4E S209 phosphorylation in WT and BBN-treated C57BL/6 mice. Representative eIF4E S209 IHC. Quantification of > 5000 cells/genotype (Normal [n = 2], 21 weeks on BBN [n = 2], P < 0.0001, t test). Scale bars: 100 μm. Data are presented as mean ± SEM. See complete unedited blots in the supplemental material.