Journal: Frontiers in Immunology
Article Title: Machine learning identifies PYGM as a macrophage polarization–linked metabolic biomarker in rectal cancer prognosis
doi: 10.3389/fimmu.2025.1639303
Figure Lengend Snippet: PYGM promotes colorectal cancer cell proliferation, migration, invasion, and M2 macrophage polarization. (A) Protein expression levels of PYGM were detected by western blotting. (B) EDU assay analysis. (C) Apoptosis rate was detected by flow cytometry. (D–F) Migration and invasion assay analysis. (G) Schematic illustration of the indirect co-culture system between colorectal cancer cells and THP-1-derived macrophages. (H) The expression levels of CD206 and CD301 were determined by flow cytometry. (I) Arg1, CD206, CD301 and IL-10 gene expression levels were detected by RT-PCR. (J) The protein levels of Arg1, CD206, CD301 and IL-10 were detected by western blotting. *p< 0.05; **p< 0.01; ***p< 0.001, compared to the corresponding groups.
Article Snippet: Western blotting used the following primary antibodies: PYGM (1:1,000; ProteinTech, 19716-1-AP), β-actin (1:4,000; ProteinTech, 66009-1-Ig), Arg1(1:1,000; ProteinTech, 16001-1-AP), CD301(1:1,000; ProteinTech, 13590-1-AP), CD206(1:1,000; ProteinTech, 32647-1-AP), IL-10(1:1,000; ProteinTech, 60269-1-Ig).
Techniques: Migration, Expressing, Western Blot, EdU Assay, Flow Cytometry, Invasion Assay, Co-Culture Assay, Derivative Assay, Gene Expression, Reverse Transcription Polymerase Chain Reaction