Figure S1. Extracellular and intracellular metabolites of commercially available and patient derived GBM cells and NHAs AB C NHA GBM NHA GBM PCA plot of Intracellular Metabolites PCA Plot of Extracellular Metabolites D

Figure S2. Total and extracted ion chromatograms of intracellular metabolites GBMNHA Total Ion Chromatogram Methionine Kynurenine Tryptophan 5-methylthioadenosine Extracted Ion Chromatogram Counts Acquisition Time (min)

Total Ion Chromatogram Extracted Ion Chromatogram GBMNHA Kynurenine 5-methylthioadenosine Counts Acquisition Time (min) Figure S3. Total and extracted ion chromatograms of extracellular metabolites

GBM Synthetic [M+H] + Counts Mass-to-Charge (m/z) Figure S4. Targeted analysis MS/MS product ion spectrum of methionine

GBM Synthetic Counts Mass-to-Charge (m/z) [M+H] + Figure S5. Targeted analysis MS/MS product ion spectrum of tryptophan

Figure S6. Targeted analysis MS/MS product ion spectrum of kynurenine GBM Synthetic [M+H] + Counts Mass-to-Charge (m/z)

GBM Synthetic Counts Mass-to-Charge (m/z) [M+H] + Figure S7. Targeted analysis MS/MS product ion spectrum of MTA

Figure S8. Total ion chromatogram and multiple reaction monitoring of different concentrations of methionine y= *x R^2 = Concentration (ng/ml) Responses Acquisition Time (min) Relative Intensity Mass-to-Charge (m/z) Counts A B C

Figure S9. Total ion chromatogram and multiple reaction monitoring of different concentrations of kynurenine Relative Intensity Acquisition time (minutes) 1000 ng 500 ng 250 ng 125 ng 63 ng 32 ng Mass-to-Charge (m/z) Counts Concentration (ng/ml) Responses AB C

Figure S10. Key metabolites in methionine pathway

Figure S11. Calibration Curve and Multiple Reaction Monitoring of SAM and SAH A C B D y= *x – R^2 = y= *x – R^2 = Concentration (ng/ml) Responses Counts Mass-to-Charge (m/z)

Figure S12. Key metabolites in tryptophan pathway Normal cells Cancer cells KMO / KYNU triggers immune surveillance MDNSC 20 OSU 35 OSU 38 OSU 34 OSU 53 OSU 96 LN18 LN229 U118 OSU 61 OSU 68 TDO2 IDO  -Actin U87 MDNSC 20 OSU 35 OSU 38 OSU 34 OSU 53 OSU 96 LN18 LN229 U118 OSU 61 OSU 68 KMO  -Actin A B

Figures U87 U118 OSU96 OSU53 OSU61OSU68 Figure S13. Insilico analysis – Connecting metabolome and proteome in GBM cells

* * * * * ** * * Figure S14. Densitometric quantification of the chemiluminescence signals