Fig. 4

Screening for maspin ligands upon EGF treatment in MCF-10A cells. A Experimental strategy used to identify maspin ligands by IP/MS. Starved MCF-10A were left untreated or treated with EGF for indicated periods of time. After IP/MS analysis, proteins exclusively found in EGF-treated samples were classified using Venn diagram, GO annotation and protein–protein interaction (PPI) networks. B–D GO enrichment analysis displaying associated biological process, molecular function and cellular component (curated by PANTHER classification system). Fisher’s exact-test was performed with an FDR < 0.05. E Reactome pathway analysis showing the most significant pathways for the identified maspin ligands. Pathways were grouped into functional clusters with ClueGO plug-in. F Interactome analysis with maspin ligands performed by Cytoscape. Generated networks for 5, 15 and 60 min were integrated into the Mentha database (Homo sapiens), which integrates protein interactions already validated by at least one experimental technique (5 min—2619 nodes; 15 min—2899 nodes; 60 min—1620 nodes). Maximal Clique Centrality (MCC) and Bottleneck topological analyses performed by Cytohubba plug-in showing the top 15 central nodes in corresponding networks. The ligands associated with folding and unfolding (red), translation (green), transcription (purple), EGFR/cell–cell adhesion (blue) and already known maspin ligands (black-bold) are highlighted