Fig. 4

The levels of HMGB1 protein are increased by USP13. A HEK293T cells were co-transfected with both Myc-HMGB1 and various doses of Flag-USP13 plasmids and WCLs were then immunoblotted. B HEK293T cells were co-transfected with both Myc-HMGB1 and Flag-USP13 plasmids, and then treated with 100 μg/mL cycloheximide (CHX) for the indicated times and WCLs were immunoblotted. *p < 0.05, **p < 0.01 using two-tailed Student’s t-test. C, D HEK293T cells were co-transfected with Myc-HMGB1 and USP13C345A/M664/739E (USP13 Mut) C and shUSP13 for 48 h (D). WCLs were immunoblotted. E HEK293T cells were transfected with Myc-HMGB1 plasmid for 48 h, and then treated with Spautin-1 for the last 18 h indicated dose and WCLs were immunoblotted