Fig. 5

Effects of HMGB1 on expression of moesin on GEnCs, anti-MPO mAb binding to GEnCs, GEnC activation and injury were mainly TLR4 dependent. Representative histogram for expression of moesin on GEnCs (a). Blockage of TLR4 and RAGE rather than TLR2 decreased expression of moesin on HMGB1-treated GEnCs (b). Among them, TLR4 had a dominate effect. Bars represent mean ± SD of repeated measurements on neutrophils for five independent experiments. Blockage of TLR4 rather than RAGE or TLR2 decreased the binding of anti-MPO mAb to GEnCs (c–g). Blockage of TLR4 rather than RAGE or TLR2 decreased the levels of LDH release (h). Blockage of TLR4 and RAGE rather than TLR2 decreased the levels of vascular barrier disruption decreased significantly (i). Bars represent mean ± SD of repeated measurements on neutrophils from five independent experiments. mAb monoclonal antibody, ANCA antineutrophil cytoplasmic antibody, GEnC glomerular endothelial cell, HMGB1 high mobility group box-1, LDH lactate dehydrogenase, MPO myeloperoxidase, MFI mean fluorescence intensity, OD optical density, RAGE receptor for advanced glycation end product, TLR Toll-like receptor