Fig. 4. PDCD4 and phospho-eIF4E control the translation of Hgf, Spp1 and Bgn.
a, Western blot showing the levels of p-4EBP1, PDCD4, eIF4A, p-MNK, p-eIF4E, eIF4G and representative HSP90 in Ptenpc−/−;Trp53pc−/−prostate cancers compared to wild-type prostates. Densitometry values normalized to the respective loading control are indicated for each band. The experiment was performed once with n = 3 mice for each group. b, Model depicting the proposed mechanism by which PDCD4 loss and phosho-eIF4E cooperate to regulate translation and modulate the tumor microenvironment. c, Fold change enrichment for the indicated mRNAs in Pten-sh TC1 determined by RNA immunoprecipitation using, from right to left, control anti-IgG, total eIF4E antibody or phospho-eIF4E antibody, followed by qRT- PCR performed (at least n = 2 independent experiments). Data are mean ± SD. Statistical analysis between all groups (ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test). d, Translation efficiency (polysomal mRNA expression/total mRNA expression) of Hgf, Spp1, Bgn, Isg15, Ccl8 and Pdgfb upon 500 nM eFT508 treatment and Pdcd4 rescue in Pten-sh TC1 prostate cancer cell line, determined by qRT- PCR (at least n = 2 independent experiments). Data are mean ± SD. Statistical analysis between all groups: (RM one-way ANOVA followed by Tukey’s multiple comparisons test).