Panel A: A flowchart depicting the screening process for identifying Golgi-nuclear localization proteins. It includes steps such as the Human Protein Atlas localization dataset, shortlisted antibodies, localization validation by siRNA interference, and bioinformatic analysis pathway enrichment. Panel B: A protein-protein interaction network showing DNA repair and membrane trafficking Golgi-nuclear localization proteins. The network is categorized by functional pathways, with DNA repair proteins in red and membrane trafficking proteins in green. Panel C: A bubble chart displaying pathway enrichment analysis of Golgi-nuclear localization proteins. The x-axis represents term enrichment, and the y-axis represents the negative logarithm of the p-value. Different pathways are represented by bubbles of varying sizes and colors. Panel D: A protein-protein interaction network showing DNA damage response proteins that localize to both the Golgi complex and nucleus. Yellow nodes indicate double-localizing proteins, and grey nodes are untested proteins. Panel E: Representative microscopy images of HeLa-K cells stained with Human Protein Atlas antibodies against Golgi-nuclear DNA damage response proteins, the Golgi marker GM130, and DNA stained with Hoechst 33342. Yellow arrowheads denote the Golgi complex. Scale bars are 10 micrometers for overview images and 5 micrometers for zoomed images.
Antibody-based analysis identifies a network of DDR proteins at the Golgi complex. (A) Schematic diagram of the screening process for the shortlisting, validation, and characterization of Golgi–nuclear localization proteins, as a strategy to identify linking pathways. (B) Experimentally based protein–protein interaction network displaying the DNA repair and membrane trafficking Golgi–nuclear localization proteins validated in this study, categorized by functional pathways. Annotations were assigned based on Gene Ontology terms and literature curation. (C) Pathway enrichment analysis of the Golgi–nuclear localization proteins identified in this study. Annotations were assigned based on Gene Ontology terms. (D) Experimentally based STRING protein–protein interaction network showing the DDR proteins identified to localize to both the Golgi complex and nucleus. DDR pathway annotations were assigned based on Gene Ontology terms and literature curation. Yellow nodes indicate double-localizing proteins, and grey nodes are filler untested proteins. (E) Representative images of HeLa-K cells stained with HPA antibodies against Golgi–nuclear DDR proteins (green), and the Golgi marker GM130 (red). DNA was stained with Hoechst 33342 (blue). Yellow arrowheads denote the Golgi complex. Scale bars, 10 μm (overview), 5 μm (zoomed images).