Fig. 4
From: Ethyl pyruvate reduces organic dust-induced airway inflammation by targeting HMGB1-RAGE signaling
EP reduces ODE-exposure induced nucleocytoplasmic translocation of HMGB1. Medium (control, 0 h) or ODE (6, 24 and 48 h post) treated cells were processed for separation of nuclear and cytoplasmic fractions and western blotting to detect HMGB1 protein. Compared to controls, ODE treated cells showed a temporal increase in HMGB1 expression (25 kD) in the (a) nuclear fraction at 6 and 48 h. Compared to vehicle (Ringer’s solution), co-treatment with EP (2.5 μM) resulted in significantly decreased levels of HMGB1 in the cytoplasm at 6 h post-treatment indicating reduction in ODE-induced nucleocytoplasmic translocation of HMGB1 (d). HMGB1 (25kD) bands were normalized over either Lamin B1 (50kD, cytoplasmic fraction, a) or β-actin (37kD, nuclear fraction, b) and percentage intensity (n = 5/group) of treatment groups relative to control were analyzed using two-way ANOVA (c and d). **p < 0.01 (* indicates difference within the OD/barn exposure groups)