Figure 3.

Differential modulation of key metabolites via Se in LPS-stimulated BMDMs indicate a Se-dependent reduction in glycolytic pathway and increased the TCA cycle. Metabolites were extracted in methanol, dried, and reconstituted in the mobile phase aqueous methanol containing 100-μM chlorpropamide as an internal standard. All samples were acquired in biological triplicate and randomized before 10 μl was acquired in LC-MS using reverse-phase UHPLC coupled to an Exactive Plus orbitrap MS. A total of 47 metabolites were profiled in biological triplicates. Individual metabolite extracted-ion chromatograms were used, with the peak area determined. All peak areas were normalized to internal standard chlorpropamide, and abundance was calculated relative to that of zero-hour zero-selenium cells (naïve cells). A, the heat map profiling of inflamed BMDMs show log2 fold change values of all 47 metabolites. All displayed log2 fold changes are represented in biological triplicates. B, the t-SNE method applied to all samples show distinct clusters separating the samples by time after LPS stimulation. BMDMs, bone marrow–derived macrophages; LPS, lipopolysaccharide; Se, selenium; TCA, tricarboxylic acid; t-SNE, t-distributed Stochastic neighbor embedding.