Metabolomics analysis-LCMS-neg

Naïve CD4⁺ T cells sorted by immunomagnetic beads from human PBMCs were cultured under Tfh cell skewing conditions for 5 days. The cells and untreated naïve CD4⁺ T cells were snap-frozen and metabolites were extracted with 80% ice-cold methanol. Liquid chromatography-mass spectrometry (LC-MS) was performed on an Ultimate 3000-Velos Pro system equipped with a binary solvent delivery manager and a sample manager, coupled with an LTQ Orbitrap Mass Spectrometer equipped with an electrospray interface (ThermoFisher Scientific). The raw data were processed with progenesis QI (Waters Corporation) and Principal component analysis (PCA) and hierarchical clustering of metabolites were analyzed.