Fig. 2
Loss of ZFHX3 compromises but does not eliminate AR’s activity in KLK3 transcription. (A-B) Detection of KLK3 protein and KLK3 mRNA by western blotting and real-time qPCR, respectively, in C4-2B and LNCaP cells cultured in the hormone-free medium for 24 h and then treated with R1881 at indicated concentrations for 24 h (A) or cultured in complete medium for 48 h (B). The transfection of siRNAs was done for 48 h before cell harvesting. (C-E) Expression plasmids of pGL3 vector control (pGL3-basic), pKLK3-Luc, and the pRL-TK reporter were transfected into C4-2B cells incubated in phenol red-free medium containing 5% charcoal-stripped FBS for 24 h (C) or cultured in complete medium (D-E). After 24 h, cells were treated with R1881 (C) or enzalutamide (D) at the indicated concentrations for 24 h. The transfection of siRNAs was done for 42 h before cell harvesting. Relative luciferase activities were then determined. The original blots can be found in Fig. S2 (Supplementary Information). ns, not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
