Extended Data Fig. 10. Asparagine Rescue and Tumor Study Controls.
A. Isotopologue abundance of aspartate from sensitive clone N and insensitive clone V treated for 4 hours with oligomycin or vehicle, then switched to media containing with universally labeled 13C-glutamine and treated with oligomycin or vehicle for an additional 4 hours (n = 3). B. Relative doubling time of clone N cells treated with 1 nM oligomycin with or without the addition of exogenous asparagine, pyruvate, or alpha-ketobutyrate (AKB) (n = 3). C. Dose response curve of clone N cells treated with oligomycin in the presence of absence of 100 µM exogenous nucleosides (n = 3). D. Plate confluence at endpoint of respiration-inhibition insensitive clone V treated with 1.5 nM oligomycin in the presence of absence of media containing all NEAAs or individual NEAAs (n = 3). E. Plate confluence at endpoint of cell lines treated with phenformin with or without 100 µM exogenous asparagine; KPC7940B and HEK-293FT 150 µM, PA-TU-8902 and MIA PaCa-2 37.5 µM (n = 3). F. Relative metabolite levels of asparagine in the serum and tumor lysate of mice bearing orthotopic KPC-7940B treated with vehicle, phenformin, asparaginase, or phenformin + asparaginase (n = 5 each). G. H&E staining of pancreas from WT C57BL/6 J treated with either vehicle or asparaginase. H-J. Dose response curves across clonal lines of the Stearoyl-CoA desaturase-1 (SCD) inhibitor CAY10566 (H, n = 3), Fatty Acid Synthase (FASN) inhibitor TVB-2640 (I, n = 3), and ATP citrate lyase inhibitor (ACLY) BMS303-141 (J, n = 3). Scale bar = 100 µm. Error bars are mean ± SD, * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001 by two-tail student’s t test (E) or one-way Anova with Tukey post hoc (B,D,F).