Figure 4
From: Deacetylation-mediated interaction of SIRT1-HMGB1 improves survival in a mouse model of endotoxemia
HMGB1 reversibly interacts with CRM1.
(A–D) HEK293T cells co-transfected with Flag-HMGB1, Flag-HMGB1K282930R, Myc-SIRT1, and/or HA-CRM1 for 48 h were incubated with LPS (100 ng/ml) or TNF-α (20 ng/ml) for 6 h and then whole-cell lysates were immunoprecipitated with an anti-Flag antibody and analyzed by Western blotting. (E) HEK293T cells were co-transfected with Flag-HMGB1, Flag-HMGB1K282930Q, Myc-SIRT1, and/or HA-CRM1 for 48 h and then whole-cell lysates were immunoprecipitated with an anti-Flag antibody and analyzed. (F) RAW 264.7 cells co-transfected with Myc-SIRT1, HA-CRM1 and Flag-HMGB1 or Flag-HMGB1K282930R for 48 h were stimulated with LPS (100 ng/ml) or TNF-α (20 ng/ml) for 24 h. Equal volumes of conditioned media were analyzed by Western blotting to detect released HMGB1.
