Figure 8.

Expression of siglec-E does not influence toll-like receptor 4 (TLR4) endocytosis in bone marrow-derived macrophages (BMDM) and bone marrow-derived dendritic cells (BMDC). (A) Wild-type (WT) and siglec-E-deficient BMDM cultured for 3 days in 1 ng/ml lipopolysaccharide (LPS) and BMDC were incubated with E. coli-GFP for 1 h a ratio of 1:10 and TLR4 and GFP expression determined by flow cytometry (B) WT and siglec-E-deficient BMDM were incubated with the indicated doses of LPS for 3 days and TLR4 expression determined by flow cytometry. (C) BMDM were incubated for 1 day without LPS or with 50 ng/ml LPS and TLR-4 and siglec-E levels determined by flow cytometry (D) Splenocytes from WT and siglec-E-deficient mice were incubated with E. coli-GFP at 1:10 ratio and gated on F4/80 positive cells and CD11c, MHCII double-positive cells and expression of TLR4 determined by flow cytometry. Two independent biological replicates were performed for each genotype.