Fig. 1

UOK262 and UOK262WT cells growth with NEAA’s in the absence (a) or presence (b) of 2 mM Gln. The cells were incubated with 100 μM of each component of NEAA separately, together or without any for 48 h. The cell density determined using the SRB assay. UOK262WT (c) and UOK262 (d) (distinct metastatic site) were subjected to up to 144 h of incubation in the presence of 2 mM Gln, 100 μM Asn, or both amino acids. Cell growth measurements were taken after 48 h, 96 h, and 144 h and expressed as a percentage of cell density at 24 h. UOK268WT (e) and UOK268 (f) (primary site) cell lines were subjected to similar analysis over time of 144 h to compare to UOK262 lines. Samples were analyzed in triplicate, and the results shown are average cell density with a standard error of the mean. Statistical analysis was performed to compare the untreated versus treated samples using unpaired, two-tailed t tests (GraphPad Prism v. 8). (*) p < 0.05; (**) p < 0.01; (***) p < 0.0001. The significance between treatment groups shown as (#) p < 0.05