Extended Data Figure 4. Primary validation that extracellular asparagine availability affects invasion and metastasis.

a, HPLC quantification of cellular free amino-acid percentages for parental 4T1 cells when the medium is supplemented with each of the NEAAs lacking in the DMEM culture medium (n = 3 replicates per cell line). b, Quantification of MDA-MB-231 Matrigel invasion rates under the same conditions as described in Fig. 3a (n = 5 invasion chambers per condition, rank-sum P < 0.001). c, HPLC quantification of cellular free aminoacid percentages for MDA-MB-231 cells when cultured in the medium conditions described in a (n = 3 replicates per cell line). d, Violet celllabelling intensity of parental 4T1 cells when grown in asparagine-lacking or -supplemented medium for the same period that the Matrigel invasion assay described in Fig. 3a was being performed (n = 3 replicates per cell line). e, Violet cell-labelling intensity of MDA-MB-231 cells when grown in asparagine-lacking or -supplemented medium for the same period that the Matrigel invasion assay described in b was being performed (n = 3 replicates per cell line).