Figure S1. Flow cytometric analysis of Toll-like receptor 4 (TLR4) -dependent CD25 expression via stimulation of nuclear factor-κB (NF-κB) reporter in Chinese hamster ovary (CHO) cells by GroEL. CHO cells (CHO/CD14/TLR4) containing the gene encoding membrane CD25 with the human E-selectin promoter, which contains NF-κB binding sites, were treated with GroEL (10 μg ml−1) or Escherichia coli lipopolysaccharide (LPS; 10 μg ml−1) for 16 h in the presence or absence of polymyxin B (50 μg ml−1). CD25 expression was analysed by flow cytometry using FITC-conjugated anti-human CD25 monoclonal antibody. The results are presented as mean fluorescence intensity.

Figure S2. Transfer of trypan blue-labeled albumin across confluent monolayers of HMEC-1 cells grown on Transwell inserts. Different number of HMEC-1 cells were seeded onto Transwell inserts and grown to confluence. Trypan blue-labeled bovine serum albumin (BSA; 100 μl, 4%) was added to HMEC-1 monolayers formed on transwell inserts for 2 h and the amount of trypan blue-labeled BSA in the lower chamber was measured using a spectrophotometer. For transmigration experiments, cells were used at a density of 5 × 104 cells per well.

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