Extended Data Fig. 5: Metastatic site influences tumor cell metabolic state.

a, Score plot of a PCA analysis of all putatively identified metabolites by LC-MS in positive ionization mode. b, Score plot of a PLS-DA analysis of putatively identified metabolites by LC-MS in negative ionization mode comparing liver and lung metastatic samples from animals with A375 human melanomas with indicated genotypes (Methods). c, VIP score plot generated based on a PLS-DA of putatively identified metabolites by LC-MS in negative ionization mode comparing liver and lung metastatic samples from animals with A375 human melanomas with indicated genotypes, showing a strong change in lactic acid, glutamic acid and a hexose (for example glucose) that is causing the separation of the groups in (b). d, Heat map of results from a suspected targeted analysis. Compounds were manually identified in the LC-MS data based on MS1 accurate mass and MS2-spectrum. The corresponding peaks were integrated, and the results normalized by 13 C2-Citric acid that was spiked to the sample as an internal standard. The compounds were selected based on the multivariate data analysis indicating a change in the glycolysis and/or TCA cycle. disease site, liver (n = 8 specimens) vs. lung (n = 10 specimens); and genotype, parental [PIP4K2C WT] and PIP4K2C KO. Statistical significance was determined using Two-tailed t-test for (d), Lactate (p = 2.36727E-05), Pyruvate (p = 0.03), Citrate (p = 0.25), a-Ketoglutarate (p = 0.18), Succinate (p = 0.004), Fumarate (p = 0.04), Malate (p = 0.04), Hexose (p = 6.72275E-06). e, Extracted ion chromatograms (left) and corresponding MS2 spectra of the putatively identified hexose (for example glucose) with an accurate mass for the [M-H]- ion of 179.5610 (top) and of lactic acid with an accurate mass for the [M-H]- ion of 89.0244 (bottom). Much lower intensities were detected in the lung samples compared to the liver samples (Right).