SPDEF and cancer: Among the top 20 genes, NR4A1, which is known to regulate cancer cell proliferation by suppressing AKT, MAPK, and NF-κB signaling pathways, was upregulated in SPDEF-overexpressing cells (Table S2).18–20 Real-time RT-PCR and western blotting assays verified that overexpressing SPDEF could increase NR4A1 mRNA and protein expression levels, while knocking down SPDEF had the opposite effect (Figs. 4b, c and S7).