Figure 10.

Effects of KYC Treatment on HMGB1 Association with TLR4 and RAGE in Lung Lysates from Hyperoxic Pups. (A) Immunoblots of TLR4 associated with HMGB1 in lung lysates from PBS- and KYC-treated hyperoxic pups. A non-depleting concentration of anti-HMGB1 antibody was used to pull-down HMGB1 from lung lysates. The pull-down was immunoblotted for TLR4 and HMGB1. These immunoblots show that the levels of TLR4 association with HMGB1 decreased in lung lysates from KYC-treated hyperoxic pups relative to the levels of TLR4 associated with HMGB1 in PBS-treated hyperoxic pups. The lower immunoblot shows that non-depleting levels of anti-HMGB1 antibody pull-down essentially equal levels of HMGB1 from each sample irrespective of how treatments modulated lung TLR4 expression (Figure 6, supplemental data Figure S6). (B) The immunoblots show how much RAGE is associated with HMGB1 in lung lysates from PBS- and KYC-treated hyperoxic pups. A non-depleting concentration of anti-HMGB1 antibody was used to pull-down HMGB1 from lung lysates. The pull-down was immunoblotted for RAGE and HMGB1. These immunoblots show that the levels of RAGE association with HMGB1 decreased in lung lysates prepared from KYC-treated hyperoxic pups relative to the levels of RAGE associated with HMGB1 in PBS-treated hyperoxic pups. The lower immunoblot shows that non-depleting levels of anti-HMGB1 antibody pull-down essentially equal levels of HMGB1 from each sample irrespective of how treatments modulate lung RAGE expression (Figure 6, supplemental data Figure S6). (C) The bar charts show the mean ± SD of relative levels of TLR4 and RAGE associated with HMGB1 in lung lysates from PBS- and KYC-treated hyperoxic pups (n=4, * = p<0.05).