Fig 1: Identification and characterization of novel palmitoylation candidates in macrophages. (A) Venn diagram comparing the S‐palmitoylated proteins identified in this study (2502 total) with the Mouse Immune SwissPalm database. The overlap indicates previously reported candidates, while 1378 proteins (orange) represent novel candidates identified in this study. (B) Venn diagram showing the contribution of each protease (trypsin, AspN, chymotrypsin, GluC) to the identification of the 1378 novel proteins. Numbers indicate proteins identified exclusively by a specific protease or shared between them. (C–E) Functional enrichment analysis of the 1378 novel candidates. Dot plots display the top significantly enriched terms for (C) GO Biological Process, (D) GO Cellular Component, and (E) UniProt Annotation Keywords. The x‐axis (Signal) represents a weighted harmonic mean of the observed/expected ratio and ‐log10 FDR, designed to balance enrichment magnitude and statistical significance. The size of the dot corresponds to the number of genes (Gene Count), and the color intensity represents the significance (‐log10 FDR). (F) Validation of novel palmitoylation candidates. ABE enrichment followed by western blotting confirmed the S‐palmitoylation of endogenous CPSF6 and GYG1 in iBMDMs. The presence of a band in the hydroxylamine‐treated sample (Hyd+) compared to the control (Hyd−) indicates specific S‐palmitoylation.