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

Figure 6. FOXA1 negatively regulates SEMA3C expression.

Figure 6

We assessed the effect of silencing of FOXA1 and POU2F1 on SEMA3C message levels. A. Knockdown of FOXA1 (siFOXA1) triggered an increase in SEMA3C levels when compared to cells treated with scrambled siRNA (siSCX); knockdown of POU2F1 (siPOU2F1) had no effect on SEMA3C expression. Knockdown of these genes was confirmed by qPCR (POU2F1) or Western blot (FOXA1; B.). In siFOXA1-treated cells where SEMA3C levels were already elevated, SEMA3C expression was further increased upon R1881 stimulation shown at both the message C. and protein D. level. Knockdown of POU2F1 had little effect on R1881 induction of SEMA3C (C, D). WCE: whole cell extract; CM: conditioned media. E. LNCaP cells were knocked down with siAR, siFOXA1, or both siAR and siFOXA1 and monitored for SEMA3C expression. Compared to LNCaP treated with scrambled siRNA, siAR-treated cells had decreased SEMA3C expression while siFOXA1 and siAR+siFOXA1-treated cells had elevated SEMA3C expression. Knockdown of AR and FOXA1 was confirmed by Western blot analysis (E). Data represent mean, ± SD; ** p < 0.01, *** p < 0.001.