Figure 2.

Temporal phosphoproteomics and proteomic analysis of THP-1 monocytes stimulated with IL-33. (A) Brief workflow depicting quantitative proteomic analysis of IL-33 stimulated THP-1 monocytes. THP-1 cells were stimulated with recombinant human IL-33 for indicated time points. Proteins were extracted from the cell lysates and subjected to in-solution trypsin digestion, followed by TMT-based chemical tagging for quantitative proteome profiling. The samples were analyzed on a Thermo Scientific™ Orbitrap Fusion™ Tribrid™ Mass Spectrometer. Mascot and SEQUEST algorithms were used for database searches. (B) Summary of the IL-33-induced phosphoproteome and total proteome. (C) Gene ontology analysis depicting the enriched biological processes for the IL-33-induced hyperphosphorylation events (D) Kinase enrichment analysis identified predicted upstream kinases for the phosphoproteins activated by IL-33. (E) Graph representing the top eight phosphatase families that were identified as being regulated by IL-33.