Mapping the intracellular HMGB1 interactome and alterations induced by Toll-like receptor 4 activation

THP1 cells were transduced with MycBioID2-HMGB1 and thereafter cultured in SILAC media for 1 week. Cells were treated with doxycycline (2 ug/ml, 48h) to induce gene expression. Thereafter, doxycycline was removed and heavy labelled cells were stimulated with LPS (100 ng/ml) for 1 hour before addition of biotin (24 hours). Light cells were used as control. Biotinylated proteins were extracted and identified with LC-MS/MS