Fig. 4: Interactomic analysis revealed that disassembly of the TRAM1-TREX1 complex promoted TREX1 mislocalization and activation of cytosolic DNA-sensing pathway during NP cell senescence.

A Schematic workflow for TREX1 interactomic analysis via Co-IP and LC-MS using a specific anti-TREX1 antibody or control antibody in normal and senescent NP cells (n = 3 biological independent samples) (Created with BioRender.com). B Top 6 enriched KEGG pathways of the differential TREX1 interactome in normal NP cells and red-marked pathways were associated with protein processing in the ER. C Chordal graph of potential proteins in KEGG pathways involved “Protein location in ER” and “Protein processing in ER”. D Endogenous forward and reverse Co-IP assay to detect the interaction of TRAM1 and TREX1 in normal and senescent NP cells (Representative blot of three independent technical experiments). E Representative agarose gel electrophoresis images of DNA fragments from the cytosolic component in P8 NP cells after co-transfection with TRAM1-overexpressing plasmid and different Flag-tagged TREX1 variant plasmids (Representative blot of four independent technical experiments). F Quantitative analysis of cytosolic genomic DNA fragments in P8 NP cells after co-transfection with the TRAM1-overexpressing plasmid and different Flag-tagged TREX1 variant plasmids (Quantification of four independent technical experiments). G Representative IF staining images of TREX1 to show subcellular localization changes of TREX1 after TRAM1 overexpression (Representative image of three independent technical experiments), bar: 20 μm. A significant p value was determined by two-tailed ANOVA (F). Mean ± SD are shown for (F). n.s. not significant.