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HMGB1 promotes localization of TLR4 coreceptors within the lipid raft. RAW264.7 macrophages were treated with 1 μg/mL HMGB1 for 60 min, and lysates were subjected to sucrose gradient fractionation. Lysates were then drawn off in 12 separate fractions and run on SDS-PAGE before immunoblot (IB) with flotillin-1 to identify raft-enriched fractions. Fractions were then probed for MD2 or CD14. Images shown are representative of at least three independent experiments.
