Figure 7.

Effect of Asn deprivation on cell death for each ASNS variant. Panel (A) replots the DMEM-Asn only data shown in Figure 6E to expand the y-axis and illustrate the differences between the control “JRS only” cells, which express no ASNS, and JRS cell lines stably expressing either the G373V or R519H variant. B, after incubation in DMEM ± Asn for the indicated time, the media containing dead/floating cells were saved in tubes and the attached cells were collected by trypsinization and added to the respective media tube. After cell collection by centrifugation, the percent dead cells were determined by trypan blue staining and cell counting. For all panels, the data are shown as the averages ± SDs of assays in triplicate. Where not shown, the SD bars are within the symbol. Each experiment was repeated at least once with different batches of cells to establish reproducibility. An asterisk indicates a p value of ≤ 0.05 compared to the control “JRS only” cells. ASNS, asparagine synthetase; JRS, Jensen rat sarcoma.