Fig. 2.

Genetic ablation or pharmacological inhibition of EZH2 leads to feedback activation of the NF-κB signaling in PCa cells. a LNCaP cells were transfected with pGL3-NFkB-Luc and pSV40-renilla plasmids for 12 h and then treated with indicated dose of EPZ6438 for additional 24 h. The luciferase activity was then measured. **P < 0.01, ***P < 0.001. b Relative mRNA expression levels of the two NF-κB downstream target genes c-Myc and Cyclin D1 in 10 μM EPZ6438 treated LNCaP cells were determined by real-time PCR assay. ***P < 0.001. c Relative mRNA expression levels of the two NF-κB downstream inflammatory cytokines IL1B and TNFA in 10 μM EPZ6438 treated LNCaP cells were determined by real-time PCR assay. ***P < 0.001. d. LNCaP cells were transfected with indicated siRNAs for 12 h, and then co-transfected with pGL3-NFkB-Luc and pSV40-renilla plasmids for 24 h. The luciferase activity was then measured. **P < 0.01, ***P < 0.001. e LNCaP cells were transfected with indicated siRNAs for 36 h, the relative mRNA expression levels of EZH2, c-Myc and Cyclin D1 were determined by real-time PCR assay. *P < 0.05, **P < 0.01, ***P < 0.001. f LNCaP cells were transfected with indicated siRNAs for 36 h, the relative mRNA expression levels of IL1B and TNFA were determined by real-time PCR assay. *P < 0.05, **P < 0.01, ***P < 0.001