The effect of DHT on cell proliferation and soft agar growth. (A) LNCaP-Flag-AR cells in phenol-red free RPMI with 5% charcoal-stripped serum were treated with DHT ranging from 0.01 nM to 10 nM with or without Wnt3A 100 ng/ml for four days in 96 well plates. The cells were then labeled with [3H]-thymidine and harvested to measure the thymidine incorporation. Cells that received no DHT treatment were used as control. The percentage of [3H]-thymidine incorporation was calculated compared with control. (B). LNCaP and LNCaP-Flag-AR cells were plated in soft agar with no treatment as control or with 100 ng/ml Wnt3A, or 10 nM DHT. After approximately 4 weeks, colonies were fixed with 10% formaldehyde in PBS. A representative field of cells was photographed for each cell type, with or without treatment using bright-field microscopy. Upper panel LNCaP or LNCaP-Flag-AR cells received no treatment, while lower panel, LNCaP-Flag-AR cells were either treated with Wnt3A or DHT.