Skip to main content
Figure 5 | Molecular Medicine

Figure 5

From: Toll-Like Receptor 4 Engagement Inhibits Adenosine 5′-Monophosphate-Activated Protein Kinase Activation through a High Mobility Group Box 1 Protein-Dependent Mechanism

Figure 5

Stimulation with LPS or overexpression of HMGB1 enhances interactions between HMGB1 and LKB1. (A) RAW 264.7 macrophages were treated with LPS (0 or 100 ng/mL) for 4 h, and then cell extracts were subjected to immunoprecipitation using antibodies to LKB1, AMPK or CAMKK. Representative Western blots and quantitative analysis show amounts of HMGB1 associated with LKB1, AMPK or CAMKK after immunoprecipitation (left panel) as well as total HMGB1, CAMKK, AMPK and LKB1 in cell extracts before (right panel) immunoprecipitation (means ± SD, n = 3, ***P < 0.001). (B) Complex formation between HMGB1 and LKB1 was determined in control MCF7-HMGB1 cells (−DOX) or after doxycycline-induced expression of HMGB1-FLAG (+DOX). Cell extracts were subjected to immunoprecipitation with ANTI-FLAG® M2 monoclonal antibody followed by Western blot analysis of LKB1. Representative blots show amounts of LKB1 after HMGB1-FLAG pull-down (left panel) as well as before immunoprecipitation (right panel). A second independent experiment provided similar results.

Back to article page