Fig. 4

NONO is a potential interaction partner for TRIM25 in GBM cells. A Venn diagram of proteins that could potentially interact with TRIM25 in LN229 and U251 cells; After excluding proteins identified in the IgG groups, a total of eight proteins were found to interact with TRIM25 in both LN229 (66 proteins) and U251 (70 proteins). B Western blotting analysis of co-IP assays performed using anti-TRIM25/anti-NONO antibody and IgG antibody, with lysates prepared from LN229 and U251 cells. C Schematic representation of full-length (FL) TRIM25 and the following four deletion mutants: amino acids 1–83 (∆RING), 84–202 (∆B-BOX), 203–409 (∆CC), and 410–630 (∆P-SPRY); CC: coiled-coil. D Western blotting analysis of co-IP assays performed on lysates prepared from HEK293 cells transfected with HA-NONO and indicated Flag-TRIM25 constructs. E Schematic representation of full-length (FL) NONO and the following three deletion mutants: amino acids 74–141 (∆RRM1), 148–229 (∆RRM2), 268–372 (∆CC); CC: coiled-coil. F Western blotting analysis of co-IP assays performed on lysates prepared from HEK293 cells transfected with Flag-TRIM25 and indicated HA-NONO constructs. G Protein-protein docking visualization (green for TRIM25, blue for NONO). a Protein-protein docking based on a hybrid algorithm of template-based modeling and ab initio free docking. b Docking interaction surfaces of TRIM25 and NONO (indicated in purple). c Microscopic diagram showing mechanism of TRIM25-NONO interaction