Immunoblot and Immunoprecipitation of Key Signaling Proteins

Immunoblot and immunoprecipitation were performed according to the standard procedures (Kim et al., 2018 (link)). The following primary antibodies were used: anti-HDAC6, TLR2, TLR4, and SIRT1 (ABclonal); anti-FcεRIβ, Lyn, GATA3, T-bet, JNK1, pJNK1^T183/Y185, Tryptase, Chymase, BECN1, MyD88, TSG101, and Calnexin (Santa Cruz Biotechnology); anti-CXCL13 (R&D Systems); anti-CD163, FoxP3, TSLP, and MIP-2 (Abcam); anti-iNOS, pBECN1^S14, COX2, ERK1/2, pERK^T204, HDAC3, NFκB, AMPKα, pAMPKα^T172, IKBα, pIKBα^S32, p38MAPK, p-p38MAPK^T180/Y182, and LC3(Cell Signaling Technology). The detailed information of primary antibodies is described in Supplemental Table S2.To isolate tissue lysates, tissue was frozen in liquid nitrogen to preserve protein structure and homogenized with RIPA buffer. After lysis, vortexing and centrifugation at 10,000 X g for 15 min at 4°C were followed. Supernatant was then obtained and used as tissue lysates for immunoblot and immunoprecipitation.