Fig. 2: XPO1 mediates TIRR nuclear export in response to DNA damage.

a Schematic representation highlighting potential nuclear export signals (yellow) in TIRR and amino acid sequence alignments of NES motifs and TIRR NES mutants. aa, amino acid. b Co-IP of whole-cell lysates from 293T cells transfected with HA-XPO1, Flag-TIRR-WT, or Flag-TIRR NES mutants, and treated with IR (8 Gy). IFC (c) and quantification (d) of cytoplasmic translocation of Flag-TIRR-WT and NES mutants in U2OS cells transfected with indicated constructs and treated with or without IR (8 Gy). Scale bar, 10 µm. Data were shown as the mean ± SD from 20 fields (>200 cells, n = 20) from three biological replicates. Two-tailed unpaired Student’s t-test; P values based on the order of appearance: 4.11E-36, 8.31E-23, 3.38E-29, 0.0042. e WB analysis of whole-cell (WCL), cytosolic (Cyto.) and nuclear (Nuc.) fractions from control or XPO1 knockout PC-3 cells post-IR (8 Gy). IFC (f) and quantification (h) of cytoplasmic TIRR in control and XPO1 knockout PC-3 cells post-IR (8 Gy). Scale bar, 10 µm. Data were shown as the mean ± SD from 20 fields (>200 cells, n = 20) from three biological replicates. Two-tailed unpaired Student’s t-test; P values based on the order of appearance: 0.0504, 2.28E-23, 6.50E-24. IFC (g) and quantification (i) of cytoplasmic translocation of TIRR in PC-3 cells pretreated with of DMSO or KPT-330 (1 μM, 12 h) post-IR (8 Gy). Scale bar, 10 µm. Data were shown as the mean ± SD from 20 fields (>200 cells, n = 20) from three biological replicates. Two-tailed unpaired Student’s t-test; P values based on the order of appearance: 0.6651, 3.83E-23, 6.39E-11. Source data are provided as a Source Data file. Similar results for (b, e) panels were obtained in three independent experiments.