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. 2017 Mar 13;7:42759. doi: 10.1038/srep42759

Figure 4. ELF5 Regulates The Expression of AR-responsive Genes.

Figure 4

(A) Quantitative PCR analysis of AR and AR responsive genes KLK3 (PSA), TMPRSS2 and FKBP5 in LNCaP cells that were transiently transfected with shScr or two different shELF5. qPCR with total RNA was performed at 72 h post transfection. Data are from two independent experiments in triplicate. (B) WB analysis of PSA and FKBP5 proteins obtained from LNCaP cells that were transiently transfected with shScr or two different shELF5. WB was performed at 72 h post transfection. (C) Quantitative PCR analysis of AR and AR responsive genes KLK3 (PSA), TMPRSS2 and ERG in VCaP cells that were transiently transfected with shScr or two different shELF5. qPCR with total RNA was performed at 72 h post transfection. Data are from two independent experiments in triplicate. (D) WB analysis of PSA and FKBP5 proteins obtained from VCaP cells that were transiently transfected with shScr or two different shELF5. WB was performed at 72 h post transfection. (E,F) PSA promoter reporter (PSA-Luc) activity assay in LNCaP cells that were transiently transfected with vector, two different shELF5, HA-ELF5 and Myc-AR, followed by EtOH or 10 nM DHT treatment for 48 h. The PSA-Luc activity assay was performed at 72 h post transfection. *compared with line 1, P < 0.05; #P < 0.05, ##P < 0.01. (G) CHIP was performed with AR-N20 antibody using material prepared from LNCaP cells that were transiently transfected with shScr or shELF5, followed by EtOH or 10 nM DHT treatment for 48 h. Normal rat IgG served as a negative control. Results were analyzed by qPCR with primers targeting potential heat shock elements in human PSA, TMPRSS2 and FKBP5. The experiment was repeated twice. *P < 0.05, **P < 0.01, ***P < 0.001. (H) CO-IP and WB analysis of AR and ELF5 proteins in total lysates obtained from LNCaP cells that were transiently transfected with shScr or shELF5, followed by EtOH or 10 nM DHT treatment for 48 h.