Cxcl1 af5403

Cells were treated as indicated and then lysed using RIPA (Beyotime Biotechnology, Shanghai, China). Protein concentration was quantified with the Bicinchoninic Acid Kit (Sigma-Aldrich) according to the manufacturer’s instructions. Equal amounts of protein (30 µg) were resolved on SDS-PAGE gel, transferred to a polyvinylidene fluoride microporous membrane (Millipore, Billerica, MA, USA) and probed with primary antibodies for β-actin (4970S, Cell Signaling Technology, CST, Boston, MA, USA), CXCL1(AF5403, Affinity Biosciences, Cincinnati, OH, USA), and β-catenin (51067-2-AP, Proteintech, Chicago, USA). The relative expression levels of proteins between groups were calculated by comparing optical densities of bands using the Gel-Pro analyzer four software (Media Cybernetics, Maryland, USA).

Cells were treated as indicated and then lysed by RIPA (Beyotime Biotechnology, Shanghai, China). Protein concentration was quantified with the Bicinchoninic Acid Kit (Sigma-Aldrich, Shanghai, China) according to the manufacturer’s instructions. Equal amounts of protein (50 μg) were loaded for SDS-PAGE electrophoresis, transferred to a polyvinylidene fluoride microporous membrane (Millipore, Billerica, MA). The signals were probed with primary antibodies and amplified by the secondary antibodies. The primary antibodies included ARG1 (DF6657, Affinity Biosciences, Cincinnati, OH, USA), iNOS (18985–1-AP, Proteintech, Rosemont, IL, USA), CXCL1(AF5403, Affinity Biosciences, Cincinnati, OH, USA), CXCR2 (20634–1-AP, Proteintech, Rosemont, IL, USA), β-actin antibody (4970, Cell Signaling Technology, Danvers, MA, USA), MMP2 (A6247, ABclonal Technology Cambridge, Boston, USA), MMP9 (10375–2-AP, Proteintech, Rosemont, IL, USA). Finally, the bands were imaged through the ECL Advance reagent (Tanon Science & Technology, Shanghai, China).