Figure 7: The effect of DPP4 inhibition on NF-κB binding (a) High glucose increased NF-κB binding and this was not reduced by the addition of linagliptin. (b) High glucose increased secreted HMGB1 after 24 hours. HMGB1 in the cell culture supernatant was quantitated using western blot. (c) DPP4 inhibition with linagliptin (30 nM) reduced recombinant HMGB1 (50 nM) induced NF-κB binding. HK2 cells were exposed to recombinant HMGB1 for 2 hours after 24 h pretreatment with linagliptin. NF-κB binding was measured with eletrophoretic mobility shift assay. N=3, *p<0.05 vs. ctrl/HMGB1. (d) Recombinant HMGB1 was not altered by pretreatment with linagliptin (30 nM) in HK2 cells. The antibody used to probe the western blot was directed to the amino terminal end of HMGB1 and should detect cleavage of HMGB1.