Figure 1.

Identification of the androgen-independent elements responsible for upregulation of L-plastin. (a) the × 400 photographs of LNCaP and LNCaP-AI cells. (b) MTT assays showed the proliferation of LNCaP and LNCaP-AI cells in steroid-depleted medium (cs-FBS). (c) The dose–response curve of LNCaP-AI and LNCaP cells proliferation over a range of DHT concentration. (d) The expressions of AR, PSA and p21 were examined by western blot analyses in LNCaP and LNCaP-AI cells. (e,f) Transcriptional activity of the L-plastin promoter was evaluated by the sequentially deletions of the androgen and oestrogen elements into the LNCaP cell line by examining the L-plastin promoter linked to Renilla luciferase activity. (g) Deletion analysis of pGL3-ΔARE1,2,3-ΔERE which is the construct with deletions of both AREs and EREs. Schematic presentation of pGL3-L-plastin-P 0.2 which encompassed nucleotides −216 to +118 in L-plastin promoter. The pGL3-basic and pGL-3-CMV were transfected as negative control and positive control. (h,i) EMSA analyses were applied to identify androgen-independent response elements in the −216 bp fragment at 3′end of L-plastin promoter. Ten pairs of distinctive, overlapping oligoes in this area for synthesized and labelled with ³²P. The arrows indicated positive band shift sites