Fig. 10
From: Ethyl pyruvate reduces organic dust-induced airway inflammation by targeting HMGB1-RAGE signaling
Treatment with EP or anti-HMGB1 neutralizing antibody decreases NF-kB p65 nuclear translocation Cells were processed for separation of nuclear and cytoplasmic fractions and western blotting to detect NF-κBp65 protein. Compared to controls, ODE treated cells showed a temporal increase in NF-κB p65 expression (68 kDa) in the (a) nuclear fraction at 15 min. Compared to vehicle (medium), co-treatment with EP and anti-HMGB1 antibody (10 μM) resulted in significant decrease in the levels of NF-κB p65 in the cytoplasm at 15 min post-treatment indicating reduction in ODE-induced NFκB p65 activation (b and d). NF-κB p65 (68 kD) bands were normalized over either Lamin B1 (50 kD, nuclear fraction, a) or β-actin (37 kD, cytoplasmic fraction, b) and percentage intensity (n = 5/group) values of treatment groups relative to control were analyzed using two-way ANOVA (c and d). * or # p < 0.05, ** or ## p < 0.01, *** or ### p < 0.001, **** or #### p < 0.0001. # indicates different from control whereas * indicates difference within the OD/barn exposure groups