Figure 12. Mislocalization of TRIM56 from the cellular nucleus to the cytoplasm is a unique molecular event occurring in senescent NP cells.

(A) Representative Western blots of USP10, USP5, TRIM56, TRIM25, and MKRN1 expression in NP cells (n = 4 biological replicates). (B) Relative mRNA expression of USP10, USP5, TRIM56, TRIM25, and MKRN1 in NP cells (n = 4 biological replicates). (C) Schematic workflow showing the IP-MS assay to identify TRIM56-interacting proteins (n = 3 biological replicates). (D) Venn diagrams showing differential ATR interactome in normal (n = 3) and senescent (n = 3) NP cells. (E) GO analysis of TRIM56 interactome showing differential pathways enriched in normal NP cells. (F) Representative Western blots showing total, cytosolic, and nuclear TRIM56 in NP cells (n = 3 biological replicates). (G) Protein sequence diagram showing the predicted NLS in TRIM56 from the NLStradamus database and PSORT. (H) Sequence conservation analysis of predicted lysine residues in multiple species. (I) Representative Western blots showing total, cytosolic, and nuclear amounts of TRIM56^WT and TRIM56^Mut in siRNA-induced, TRIM56-deficient P2 NP cells after transfection with Myc-tagged plasmids. Data are presented as the mean ± SEM. At least 3 independent experiments were performed. NS, not significant; unpaired Student’s t test (B).