Figure 6.

Cell viability of ex vivo CRPC xenograft after siRNA knockdown of ASNS. MTT cell growth assays. PCA 180-30 cells (0.7 million) from primary cultures (see Materials and Methods) were seeded into each well of a six-well plate. After 48 hours of incubation, the cells were transfected with control siRNA (si-control) or si-ASNS. At 24 hours after transfection, the cells were trypsinized, and 4000 cells in 100 μL of complete medium were seeded into each well of a 96-well plate. These cells were continually incubated for 48 hours, followed by no treatment (A) and l-asparaginase treatment (B). The treatments and MTT assays were performed as described in the legend to Figure 4. C: Western blot analysis to evaluate the performance of si-ASNS in PCA 180-30 cells. The cells were treated in the same manner as those described in the legend to Figure 4, and the Western blot analysis was performed as described in the legend to Figure 4C.