Fig. 3

Mechanism dissection how AR suppresses lncTCFL5-2 expression. A SW839 cell were virally transduced with sh-AR1, sh-AR2 and cells were exposed to hypoxia or normoxia for 2 days. qPCR analysis of the expression of lncTCFL5-2. B OSRC-2 cells were virally transduced with oe-AR and cells exposed to hypoxia or normoxia for 2 days, qPCR analysis of the expression of lncTCFL5-2. C SW839 cells were treated with Enzalutamide (Enz) 10 µM and then cells exposed to hypoxia or normoxia for 2 days, qPCR analysis of the expression of lncTCFL5-2. D The interaction between AR and lncTCFL5-2 in both SW839 and OSRC-2 cells under normoxia and hypoxia condition were detected by RIP. E Bioinformatic analysis of potential AR binding sites in lncTCFL5-2 promoter. F Lysates of OSRC-2 cells were subjected to ChIP assay. ChIP products were amplified by PCR reaction. G, H SW838 and OSRC-2 cells were co-transfected with the indicated reporter constructs and Renilla luciferase plasmid. 48 h after transfection, reporter activity was then measured and plotted after normalizing against the Renilla luciferase activity (mean ± SD).. (pLKO: pLKO.1-puro plasmid, sh-AR1/2: The plasmid encoding AR-shRNA1/2, oe-AR: The plasmid encoding AR sequence, pWPI: Addgene plasmid 12,254, ARE:AR Element)