Figure 3.

BTK shRNA and BTK-(C)specific siRNAs knock down decrease cell survival in prostate cancer cell lines. LNCaP (A) and DU145 (B) were transfected with shRNA and co-transfected with GFP to mark transfected cells. Transfected cells were counted at 24h and 72h and the 72h to 24h ratio was calculated and expressed as % of the control. LNCaP (C) and DU145 (D) were transfected with BTK-C specific siRNA or non-targeting siRNA. Co-transfection with a GFP expressing plasmid marks transfected cells. Transfected cells were counted and the 96h to 24h ratio was calculated and expressed as % of the control. (E) DU145 and NAMALWA cells were transfected with BTK-C specific siRNA and control siRNA for 48h. The cell lysates were prepared for immunoblotting. GAPDH is a loading control; the results show BTK-C siRNA just decreases the BTK-C protein and not BTK-A protein in NAMALWA cells. LNCaP (F) and DU145 (G) were transfected with BTK-C specific siRNA or non-targeting siRNA as a control for 48h. Increased cleaved caspase-3 was detected compared with control. Apoptotic cells for each treatment were calculated as fold increase in Caspase-3 positive cells of control. Mean of triplicate assays ± SD. Student t-test, *p < 0.05.