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. 2021 Jan 13;4(1):327–337. doi: 10.1021/acsptsci.0c00196

Figure 1.

Figure 1

(A) UCSC genome browser track encompassing the ASNS locus representing the location of the sequences targeted by the two sgRNAs employed to generate ASNS KO cells. Red boxes represent exons encoding for the asparagine synthetase domain. (B) Western blot analysis of A2058 cells bearing doxycycline inducible sgASNS constructs upon doxycycline treatment (lane 1 and 2). Single cell clones derived from transient expression of sgASNS (lane 3–10). Vinculin is used as a loading control. (C) Relative growth (Log10 cell number values as measured by Incucyte – y-axis) for A2058 wild type and ASNS KO clones. The x-axis represent the hours elapsed after NEAA deprivation from cell culture media. (D–E) Relative abundance of the indicated amino acids in A2058 WT and KO cells at 6 h (D) and 24 h (E) removal of NEAA. (F) Relative viability (measured by Cell titer Glo – y-axis) of A2058 WT and KO cells upon increasing concentration (x-axis) of l-asparaginase.