Fig. 1.

Identification of AREs responsible for Pp-dependent transcription in epididymal cells. A, Schematic diagram of a wild-type (Wt) Pp construct and ARE-mutant derivatives (mAREs). All constructs harbor 0.3 or 0.6 kb of 5′-flanking sequence and have the Renilla luciferase gene downstream (not shown). B–D, Luciferase analysis performed using epididymal cells incubated with or without the synthetic testosterone R1881 (T) and transiently transfected with the constructs shown in A (100 ng) and a simian virus 40 promoter-driven firefly luciferase plasmid PGL3-E-V (50 ng), the latter of which serves as an internal control for normalization. Some cells were also cotransfected with an AR expression vector (100 ng). Shown are average values ± se from three experiments done in triplicate (values are relative to cells transfected with the empty Renilla luciferase expression vector, which was given a value of 1). E, Real-time PCR analysis of total cellular RNA from adult wild-type and SPARKI mice epididymides was done using at least four independent samples that were normalized to the level of L19 mRNA, which encodes a ribosomal protein. mRNA levels in control mice were given a value of 1. Average values ± se are shown. TSS, Transcription start site; m, mutant. An asterisk indicates statistically significant differences from the control (P ≤ 0.05).