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. Author manuscript; available in PMC: 2013 Jul 16.

Published in final edited form as: Prostate. 2010 Jun 15;70(9):934–951. doi: 10.1002/pros.21128

Fig. 6 — TS1 and TS2 share similar biological function on −286PB promoter. (A) and (B), Similar transcription factors bind to the TS1 and TS2 in LNCaP cells. The radiolabeled PB –257 to –232 bp probe containing the TS1 (A) and PB –127 to –102 bp probe containing the TS2 (B) were incubated with 10 μg of the LNCaP nuclear extracts in the absence or presence of 200×, 100×, 50×, 25×, 12.5× and 6.25× unlabeled TS1 or TS2 competitors. (C), Schematic representation of the wild type – 286PBLuc, TS1/TS1, TS2/TS2, and TS2/TS1 reporter constructs. (D), LNCaP cells were transiently transfected with wild type –286PBLuc, TS1/TS1, TS2/TS2, and TS2/TS1 reporter constructs together with the AR and Renilla luciferase expression vectors and relative Luc activities were determined in the absence or presence of 10⁻⁸ M DHT as described in Materials and Methods. The reporter gene activities were measured in at least three determinations ± SD, and all experiments were adjusted for transfection efficiency. The luciferase activities are expressed as relative luciferase light units/min/μg protein.

Fig. 6 — TS1 and TS2 share similar biological function on −286PB promoter. (A) and (B), Similar transcription factors bind to the TS1 and TS2 in LNCaP cells. The radiolabeled PB –257 to –232 bp probe containing the TS1 (A) and PB –127 to –102 bp probe containing the TS2 (B) were incubated with 10 μg of the LNCaP nuclear extracts in the absence or presence of 200×, 100×, 50×, 25×, 12.5× and 6.25× unlabeled TS1 or TS2 competitors. (C), Schematic representation of the wild type – 286PBLuc, TS1/TS1, TS2/TS2, and TS2/TS1 reporter constructs. (D), LNCaP cells were transiently transfected with wild type –286PBLuc, TS1/TS1, TS2/TS2, and TS2/TS1 reporter constructs together with the AR and Renilla luciferase expression vectors and relative Luc activities were determined in the absence or presence of 10⁻⁸ M DHT as described in Materials and Methods. The reporter gene activities were measured in at least three determinations ± SD, and all experiments were adjusted for transfection efficiency. The luciferase activities are expressed as relative luciferase light units/min/μg protein.