Ab24157

Protein samples of the cells were extracted by lysing the cells with radio immunoprecipitation assay lysis buffer (Beyotime, Shanghai, China). The protein samples were quantified using the BCA Protein Assay Kit (Beyotime) and the same amount of samples were loaded for sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein bands on the gel were transferred to a polyvinylidene fluoride membrane and blocked in 5% skim milk for 2 h at room temperature and then the blot was incubated in the specific primary antibodies for TSLP (ab47943), skeletal muscle actin (α-SMA, ab5694), collagen I (ab6308), MAPK7 (ab40809), extracellular signal-regulated kinase 1 (ERK1, ab180163), phospho-ERK1 (p-ERK1, ab24157), p38 (ab7952), p-p38 (ab178867), c-Jun N-terminal kinase 1 (JNK1, ab199380), and p-JNK1 (ab47337), purchased from Abcam (Cambridge, UK), overnight at 4°C. β-actin (ab189073) was used as an endogenous reference. After washing, the blot was incubated in horseradish peroxidase-conjugated secondary antibodies for 1 h at room temperature. Positive signals were developed using the ECL Plus Western Blotting Substrate (Thermo Scientific). Band densities for each sample were analyzed using ImageJ 1.49 software (National Institutes of Health, Bethesda, MD).