Fig. 2.

Binding domain study between HMGB1 and USP13. A HMGB1 wild type (WT), A, B boxes (upper panel). HEK293T cells were co-transfected with either Myc-HMGB1 A or B box and Flag-USP13 plasmid (lower panel). Immunoprecipitations (IPs) using anti-Flag and anti-Myc antibodies were done. ns: non-specific. WCLs were immunoprecipitated (IP) and immunoblotted (IB) with indicated antibodies. B USP13 WT, USP13 ZnF, and USP domains (upper panel). HEK293T cells were co-transfected with both Myc-HMGB1 and Flag-USP13 WT or USP13 ZnF or USP13 USP domain plasmids (lower panel). WCLs were immunoprecipitated (IP) and immunoblotted (IB) with indicated antibodies. C HEK293T cells were transfected with Flag-USP13 WT, ZnF, and USP domain plasmids alone, and immunoprecipitated with anti-HMGB1 Ab to confirm the binding of the domain of USP13