A11504

On day 14 after tumor implantation, lung tissues were collected, homogenized, and lysed in RIPA buffer to extract total protein. Protein concentration was measured using a BCA protein assay kit (P0012, Beyotime). Absorbance was measured, and protein concentration was determined using a standard curve. The 30 μg protein samples were subjected to SDS-PAGE and subsequently transferred to PVDF membranes (G6015, Servicebio) before blocking (GF1815, Genefist Life Science CO., LTD) and incubating with primary antibodies.^{22 (link)}
Primary antibodies included anti-S100A8 antibody (1:1000; A1688, Abclonal), anti-S100A9 antibody (1:1000; A9842, Abclonal), anti-matrix metalloproteinase 9 (MMP9) antibody (1:1000; ab76003, Abcam), anti-lysyl oxidase (LOX) antibody (1:1000; A11504, Abclonal), anti-signal transducer and activator of Transcription 3 (STAT3) antibody (1:2000; A1192, Abclonal), anti-p-STAT3 antibody (1:2000; AP0705, Abclonal), and anti-β-actin antibody (1:2000; AC006, Abclonal). The secondary antibody (1:5000; GB23303, Servicebio) was added and incubated at room temperature for 1 hour. Proteins were then visualized using an ECL luminescent reagent (PM00021, Abclonal), and the densitometric analysis of the protein bands was analyzed using Image J software.