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. 2016 Dec 25;8(6):9617–9633. doi: 10.18632/oncotarget.14168

Figure 2. SEMA3C is an androgen receptor-regulated gene.

Figure 2

LNCaP were treated with increasing concentrations (0 to 5 nM) of the synthetic androgen, R1881 A., and increasing concentrations (0 to 10 nM) of dihydrotestosterone (DHT; B.) followed by detection of SEMA3C mRNA levels by qPCR; relative quantities (RQ) are presented. R1881-stimulated LNCaP cells were treated with increasing concentrations (6.25, 12.5, 25, and 50 μM) of Enzalutamide (MDV3100; C.) or of the AR DBD inhibitor VPC-14449 (14449; D.) followed by SEMA3C mRNA level detection by qPCR. AR-negative PCa lines PC-3 and DU 145 did not upregulate SEMA3C in response to R1881 E & F. PC-3 and LNCaP cells were transfected with mock or AR overexpression plasmids followed by qPCR for SEMA3C G & H. LNCaP cells were transfected with siRNA directed against AR (siAR) or scrambled siRNA (siSCX) followed by qPCR for SEMA3C I. LNCaP cells were treated with R1881 (5 nM) in the presence of siAR followed by qPCR for SEMA3C J. LNCaP were treated with PI3K inhibitor LY294002 at the indicated concentrations or DMSO and monitored for SEMA3C message expression K. LNCaP were treated with LY294002 (40 μM) in the presence of siRNA for AR (siAR) or GATA2 (siGATA2) followed by qPCR for SEMA3C L. 22Rv1 were treated with PTEN inhibitor bpV(HOpic) at the indicated concentrations or DMSO and monitored for SEMA3C message expression M. Data represent mean, ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.