Selective growth inhibition of ASNS-negative cancer cells by WT, Q59L, and Q59F l-ASP. (A) Western blot analysis of ASNS levels in OVCAR-8 cells after 48 hours transfection with ASNS siRNA (siASNS) or negative control siRNA (siNeg). β-actin was used as a loading control. (B) WT and Q59L l-ASP concentration-activity curves. The OVCAR-8 cell line was transfected with negative control siRNA (siNeg) or ASNS siRNA (siASNS) for 48 hours, then treated with a range of l-ASP concentrations for 48 hours, and finally assayed with CellTiter-Blue. WT, Q59L, and Q59F l-ASP concentration-activity curves were determined in the (C) Sup-B15 and (D) RS4;11 leukemia cell lines by CellTiter-Blue assay. (E) Western blot analysis of ASNS levels in Sup-B15 and RS4;11 cells treated with an EC50 dose of l-ASP. No treatment was used as a primary control, and MOLT-4 cells were included as a secondary control.
