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. 2016 Apr 25;113(19):5263–5268. doi: 10.1073/pnas.1524291113

Fig. 5.

Fig. 5.

Importin 8 is elevated in AML and is an eIF4E mRNA export target. (A) Importin 8 protein levels in patients with M4/M5 AML by Western blot. Numbers refer to different patients, and normal (Norm) refers to specimens from two different healthy volunteers. Actin is a loading control. (B) Nuclear import assays using wild-type eIF4E-GST (red) as a function of treatment with RTP at 50 μM, a clinically achievable concentration (9). DAPI is in blue. Confocal micrographs represent single sections through the plane of the cells with a magnification of 63× and no additional zoom. Quantification is shown in Fig. S1H. (C) Fold mRNA enrichment relative to nuclear input in eIF4E IPs from U2OS nuclear lysates as indicated. (D, Left) RNA export assay showing the mRNA c/n for importin 8 mRNAs. (D, Right) Total mRNA levels for Ubc, importin 8, and Mcl-1 do not change upon eIF4E overexpression. Data are the mean ± SD. **P < 0.01; ***P < 0.001. ns, not significant. (E) Western blot analysis indicates that eIF4E overexpression increases importin 8 levels. (F) Model for eIF4E import cycle. Details are provided in the main text.