<b>Protein-Protein interaction (PPI) network of differentially acetylated proteins</b><b> by Aspirin during differentiation of THP-1 cell towards macrophage</b>

The human THP-1 monocytic cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin/streptomycin (Gibco) at 37°C under 5% CO₂ in two groups. The control group received 0.1% DMSO, while the treatment group was exposed to 300 μg/ml Aspirin (Sigma-Aldrich, A2093) for 3 hours. Both groups were then differentiated into macrophages using 100 nM phorbol 12-myristate 13-acetate (PMA) for 48 hours. Following differentiation, the quantitative acetyl-proteomics analysis was done to map Aspirin-driven lysine acetylome remodeling during macrophage differentiation in THP-1 monocytes. The protein-protein interaction (PPI) networks were generated using STRING (H. sapiens; confidence score &gt; 0.7) and visualized in Cytoscape 3.2.1. to elucidate how Aspirin-driven acetylated proteins functionally coordinate within cellular systems. The PPI network was further analyzed to identify densely interconnected functional clusters/modules using topological clustering algorithms. This analysis revealed 5 key functional clusters (Cluster 1 to 5) significantly affected by Aspirin-driven acetylation.