Fig. 4

EZH2 is a critical for CREB to induce NE phenotypes. a H3K27me3 level (left), as well as expression of EZH2 and NE markers (right), are up-regulated in PC3 and LNCaP cells overexpressing EZH2 cDNA. b RT-qPCR measures the expression of NE markers CHGA, CHGB in NE1.3 cells treated with EZH2 inhibitors GSK126 (5 μM) or DZNEP (5 μM) for 48 h (left). Western blotting confirmed the reduction of H3K27me3 in NE1.3 cells treated with these EZH2 inhibitors (right). Error bars in PCR results represent standard deviation (s.d.) of triplicate experiments. c RT-qPCR result shows that EZH2 knockdown by a validated shRNA downregulates the expression of NE markers CHGA and CHGB in NE1.3 cells. d PC3 cells carrying Dox-inducible EZH2 cDNA were infected with scramble shRNA or shEZH2 lentiviruses, then treated without or with 1 μg ml⁻¹ of Dox for 72 h. e RT-qPCR analysis of the changes in NE marker expression in LNCaP cells after treating with 10 μM forskolin (FSK) with or without 5 μM EZH2 inhibitor GSK126 for 24 h. f LNCaP cells expressing either control shRNA or shEZH2 were treated with 10 μM FSK for 5 h. CHGA, CHGB, and ENO2 levels were measured by RT-qPCR, using GAPDH as the loading control