Extended Data Fig. 9: AKT inhibition increases PDCD4 levels and cooperates with eFT508 to reduce HGF, SPP1 and BGN protein levels in Pten−/−;Trp53−/− prostate cancer cells.

a, Western blot analysis showing the protein levels of PDCD4 and phospho-S6 in Pten−/−;Trp53−/− (RapidCap) and PC3 prostate cancer cell line upon treatment with the indicated concentration of ipatasertib. Densitometry values normalized to the loading control are indicated at the bottom for each band. The experiment was repeated two independent times with similar results. b, Western blot analysis showing the protein levels of PDCD4, p-S6, p-eIF4E, p-4EBP1, HSP90 (upper panel) and HGF, SPP1, BGN and representative HSP90 (lower panel) in Pten−/−;Trp53−/− (RapidCap) murine prostate cancer cell line upon treatment with vehicle, 500 nM eFT508, 500 nM ipatasertib or the dual treatment. Densitometry values normalized to the loading control are indicated at the bottom for each band. The experiment was repeated two independent times with similar results. c, Western blot analysis showing the protein levels of PDCD4, p-S6, p-eIF4E, p-4EBP1, HSP90 (upper panel) and HGF, SPP1, BGN and representative HSP90 (lower panel) in PC3 human prostate cancer cell line upon treatment with vehicle, 500 nM eFT508, 500 nM ipatasertib or the dual treatment. Densitometry values normalized to the loading control are indicated for each band. The experiment was repeated two independent times with similar results. d, Transwell migration assay performed with bone marrow-derived MDSCs tested for the capability to migrate toward PCa medium conditioned with the indicated treatments. n = 3 biological replicates. The experiment was repeated two independent times with similar results. Data are mean ± SD. Statistical analysis between all groups (ordinary one-way ANOVA followed by Tukey’s multiple comparisons test). e, Representative FACS plot of the gating strategy for the quantification of CD45+/CD11b+/Ly6Ghigh/Ly6Clow (PMN-MDSCs) (left) and CD45+/CD3+/CD8+ cells (right) in Ptenpc−/−;Trp53pc−/− prostate tumors.