Py216gsk3β

Murine lung epithelial (MLE12) cells (ATCC, Manassas, VA) were cultured with HITES medium containing 10% fetal bovine serum (FBS). RAW264 cells were cultured with DMEM medium containing 10% FBS. The cells were cultured at 37°C in a 5% CO2 incubator. V5 antibody, mammalian expressional plasmid pcDNA3.1 TOPO /His-V5, Escherichia coli Top 10 competent cells, and phospho-serine antibody were from Invitrogen (Carlsbad, CA, USA). FAK, pY576/577-FAK, Flag tag (9A3), GSK3β, and pY216GSK3β antibodies were from Cell Signaling Technology (Danvers, MA, USA). Recombinant mouse IL-33 protein, KC, and mouse IL-6 ELISA kits were from R&D Systems (Minneapolis, MN). ST2L antibody was from Abcam (Cambridge, MA). FITC conjugated ST2L (FITC-ST2L) antibody (DJ8) and a Rat FITC conjugated IgG1 isotope control antibody were from MD Bioproducts (St. Paul, MN). GSK3β shRNA and scrambled shRNA were from Sigma Aldrich (St. Louis, MO, USA). FAK inhibitor and TWS119 were from Cayman Chemical Company (Ann Arbor, MI). Immunobilized protein A/G beads were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All materials used in these experiments were of the highest grade commercially available.

Mouse lung epithelial cells (MLE12) and Human monocyte-like cell line (THP1) were purchased from American Type Culture Collection (ATCC, Manassas, VA). Recombinant human IL-37 was from R&D Systems (Minneapolis, MN, USA). Cycloheximide, leupeptin, and LPS were purchased from Sigma Aldrich (St Louis, MO, USA). MG-132 was purchased from EMD Chemicals (Gibbstown, NJ, USA). Immobilized protein A/G beads and control IgG and anti-Sigirr (for mouse, rat, and human origin) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-USP13 was purchased from Protein Tech (Chicago, IL, USA) and Bethyl Laboratories (Montgomery, TX, USA). Antibodies against HA tag, GSK3β, pY216GSK3β, phospho-threonine, and ubiquitin were purchased from Cell Signaling (Beverly, MA, USA). Anti-V5, mammalian expression plasmid pcDNA3.1/His-V5-topo, Escherichia coli Top10 competent cells were purchased from Invitrogen and Thermo life technology (Gaithersburg, Maryland, USA). Horseradish peroxidase-conjugated secondary antibodies were purchased from Bio-Rad Laboratories (Hercules, CA, USA). All commercially available materials used were of the highest quality.