Fig. 2: Untargeted lipidomic analysis of lymph, plasma and their EVs.

Matched mice lymph and plasma, and EVs isolated from those two respective biological fluids, were analyzed by LC-QTOF-MS. Principal component analysis (PCA) plots for (a) plasma in blue and lymph in red and for (b) plasma EVs in blue and lymph EVs in red (PC 1, PC 2 and PC 3 refer to Principal Components 1, 2 and 3 on the Y, X and Z axes, respectively). c Volcano plot of untargeted lipidomics in lymph and plasma. The x axis represents the fold changes (in log2) of MS signal intensities values in lymph vs. plasma and the Y-axis represents the p-values (in –log10). Selection criteria were established with corrected p < 0.05 (horizontal blue line) and FC > 10 or < 0.1 (vertical blue lines), and with less stringent criteria of selection using FC > 1.3 or < 0.77 (vertical red lines). d Volcano plot of untargeted lipidomics in lymph EVs and plasma EVs. Selection criteria were established with corrected p < 0.05 (horizontal red line) and FC > 1.2 or < 0.83 (vertical red lines). Features that were discriminating lymph and plasma (or lymph EVs and plasma EVs) were annotated and validated using MS/MS analysis (color symbols). n = 3 (each n is a pool of 2–4 mice). PC: Principal Component; Car: acylcarnitines; CE: cholesteryl esters; FFA: free fatty acid; HexCer: hexosyl Ceramides; LPC: lysoglycerophosphatidylcholine; LPE: lysoglycerophosphatidylethanolamine; LPCO-: ether-linked lysoglycerophosphatidylcholine; PC: diacylglycerophosphatidylcholine; PE: diacylglycerophosphatidylethanolamine; PCO-: ether diacylglycerophosphatidylcholine; DG: diacylglycerols; SM: sphyngomyelins; TG: triacylglycerols.