Extended Data Fig. 7: DON and ASNase co-treatment in 779E cells, glutamine quantification, and correlation analyses for DON target genes in PDOs.
(a, b) Relative number of 779E cells treated with the indicated doses of DON in combination with ASNase (0.5 U/ml; +ASNase) or without (−ASNase) for 24 h. Cells were stained with crystal violet and representative images are shown. Quantification of crystal violet staining is shown relative to −ASNase 0 μM DON condition and is representative of two independent experiments. The coefficient of drug interaction (CI) was calculated for each DON concentration (shown in graph). Data are presented as mean ± SEM of n = 3 replicate wells. 0.25 mM, P < 0.0001. 0.5 mM, P = 0.0004. 1 mM, P = 0.00017. (c) Determination of the glutamine concentration in cell culture media incubated with or without 0.5 U/mL ASNase for 24 hours. Data are presented as mean ± SEM of n = 3 replicate samples. (d–k) Correlation analysis between normalized gene expression of the indicated enzymes targeted by DON and DON viability ratio for the six PDOs assessed. Statistical significance was calculated using unpaired two-tailed Student’s t test (b, c) or two-tailed Spearman correlation coefficient (d–k). ***P < 0.001, ****P < 0.0001, ns = not significant.
