Secreted CXCL8 or IL-37 protein levels in cell media were measured using CXCL8 or IL-37 DuoSet ELISA Development Systems, respectively (R&D Systems, Minneapolis, MN, USA) at 23 °C, and protein levels were calculated using a recombinant CXCL8 or IL-37 protein standard curve according to the manufacturer’s instructions.
To measure the intracellular production of IL-37, protein extraction from cells was performed in radio-immunoprecipitation assay (RIPA) buffer (Sigma-Aldrich, St Louis, MO, USA) containing a 1:100 protease inhibitor cocktail (catalogue no: P8340, Sigma-Aldrich) [6 (link)]. The detection of intracellular IL-37 from cell lysate was performed using a human IL-37 ELISA Kit (Adipogen, Life Sciences, Liestal, Switzerland) due to higher sensitivity (10 pg/ml). Protein levels were calculated using a recombinant IL-37 protein standard curve according to the manufacturer’s instructions.
To measure the intracellular production of IL-37, protein extraction from cells was performed in radio-immunoprecipitation assay (RIPA) buffer (Sigma-Aldrich, St Louis, MO, USA) containing a 1:100 protease inhibitor cocktail (catalogue no: P8340, Sigma-Aldrich) [6 (link)]. The detection of intracellular IL-37 from cell lysate was performed using a human IL-37 ELISA Kit (Adipogen, Life Sciences, Liestal, Switzerland) due to higher sensitivity (10 pg/ml). Protein levels were calculated using a recombinant IL-37 protein standard curve according to the manufacturer’s instructions.
