Fig. 3: DPYSL5 promotes stemness through the activation of the PRC2 complex.

a Gene set enrichment analysis (GSEA) reveals that DPYSL5 overexpression (DPYSL5 OE) in LNCaP cells promotes the expression of genes commonly upregulated in NEPC (NEPC-signature) and in embryonic stem cells. Additionally, upregulation of EZH2 targets and genes related to invasiveness is observed in response to DPYSL5 overexpression. (p-value > 0.001 and normalized enrichment scores (NES) > 2 were used as cut offs). b DPYSL5 overexpression promotes neuron-like morphology in CAM tumors in C42B cells when compared to control (CTRL) cells. Brown: DPYSL5 immunohistochemical staining. c DPYSL5 overexpression leads to a significant increase in the number of neurite branch points and neurite length in LNCaP cells. p-values are shown as asterisks (*p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001). d DPYSL5 overexpression increases mRNA expression of NE-markers NSE, CGA and ASCL1 in LNCaP cells based on qPCR analysis. e Additionally, protein expression of Snail increases in LNCaP cells in response to DPYSL5 overexpression. f Spheroid formation of LNCaP CTRL cells and DPYSL5 overexpression cells. DPYSL5 overexpression promotes invasiveness and spheroid growth based on analysis using IncuCyte live-cell imaging system and the associated software. g DPYSL5 overexpression results in the upregulation of genes related to progenitor cells and stemness, while H3K27 trimethylation-associated genes are downregulated. h DPYSL5 overexpression leads to the upregulation of central stem cell markers Nanog and SOX2 in LNCaP cells. i DPYSL5 overexpression increases protein levels of epigenetic regulators EZH2, EZH1 and SUZ12, and also of trimethylated H3K27 (H3K27me3).