Figure 4.

A. Immunoblots of protein lysates of xenograft tumors derived from LHSR-AR cells transduced with doxycycline-inducible constructs (in the pTRIPz vector) for the expression of GFP or NEK6 in male mice, with doxycycline maintained in the diet at time of harvest (+dox) or with doxycycline diet removed 7 d prior to harvest (−dox). Tumors were harvested from non-castrated mice (day 0) or 2 or 5 d after castration. Short and long exposures of the NEK6 immunoblot are shown. B. Gene Set Enrichment Analysis performed on the gene expression signature mediated by NEK6 overexpression at d 5 after castration (GSEA pre-ranked based on ratio of classes to tumors without NEK6 overexpression). The top two curated gene sets (C2) from the molecular signatures database (MSigDB) correlated with the NEK6 signature are shown. C. Left: Genes downregulated in the control (−dox) tumors after castration (fold change <−1.5, signal-to-noise <−1 in the comparison of tumors harvested at d 5 and day 2) were plotted against the NEK6 signature at d 5 using GSEA pre-ranked. Right: Venn diagram of genes downregulated in control tumors after castration intersected with genes upregulated by NEK6 at day 5 (fold change >1.5, signal-to-noise >1 in the comparison of +dox to −dox tumors). The GO terms most highly enriched in this overlap are cytokine-mediated and type I interferon signaling pathways (p=2×10⁻⁵).