Fig. 4: Proteomic analysis reveals stable cys-modifications in M1 BMDMs.

a Experimental design for mitochondrial-enriched proteomics with a focus on non-reversible modifications. b Volcano plot showing significant proteins with reduction resistant cysteine modifications with FC < 1 in stimulated WT BMDMs from male and female mice vs ctrl (mixed sex in all groups) (n = 4 mice). Data were corrected for total protein abundance. In blue are indicated proteins belonging to interactome of DLD together with DLD itself. c Total GO enrichment of proteins from (b) (protein number = 149). d Proteomics network of protein-protein interactions, depicting proteins as nodes and interactions as edges. e Node color is associated with the assigned network cluster. The most enriched GO term associated with each cluster is indicated. f Venn diagram indicating number of proteins belonging to interactome of DLD out of total (4b); g Enrichment analysis of subgroup of proteins from 4b-f (protein number = 31). h Volcano plot showing significant proteins with reduction resistant cysteine modifications with FC < 1 in stimulated Nos2 ^−/− BMDMs from male and female mice and treated with AS vs stimulated only (mixed sex in all groups). Data were corrected for total protein abundance (n = 4 mice). i Venn diagram indicating groups of proteins from (b) and (h) with respective overlap indicating possible protein modifications driven by endogenous HNO. Pathway enrichment analysis (GO molecular function) ( j) and network analysis (k) of merged list from (i). Data shown are representative of two or more independent experiments. Source data are provided as a Source Data file.