Figure 6.

ASNS cDNA constructs encoding WT, N80S, and S480F were independently stably expressed in an ASNS-null background JRS cell line. (A) Co-expression of GFP with ASNS cDNA constructs permits evaluation of retroviral transduction efficiency in generation of the JRS model system. ASNS mRNA (B), as determined by qRT-PCR, and relative ASNS protein abundance (C) evaluated by immunoblotting for each of the generated JRS cell lines. (D) Cellular proliferation was determined by cell counting of the JRS cell lines in the presence or absence of Asn in the media for 0, 24, 48, and 72 h. The data are the averages ± standard deviations of three determinations, and an asterisk indicates that the DMEM—Asn value is statistically different from the Asn-containing condition with a p-value of ≤0.05.