Fig. 5.

USP13 regulates subcellular localization of HMGB1. A, C HEK293T cells were co-transfected with both GFP-HMGB1 and Flag-USP13 or empty plasmids for 48 h. Cells were treated with 10 μM Spautin-1 for 2 h or 20 ng/mL leptomycin B (LMB) for 20 h and further treated with 50 μM H₂O₂ for 4 h. HMGB1 was observed after immunofluorescent staining. Over 100 cells were counted for cytoplasmic GFP-HMGB1 ( +) cells. Scale bar: 10 μm. n.s. no significant, N = 3. **p < 0.01 and ***p < 0.001 using Tukey’s honestly significant difference post-hoc test for multiple comparisons. B HEK293T cells were co-transfected with Myc-HMGB1 and Flag-USP13 plasmids for 48 h and treated with 10 μM Spautin-1 for the last 18 h and WCLs were immunoprecipitated (IP) with anti-Myc antibody and immunoblotted (IB) with anti-CRM1 antibody. Data represents one of two similar independent experiments in B