Figure 2.

BCM decreases cell surface expression of TLR4. (a) Western blot showing the total TLR4 expression in BMDM whole-cell lysate, which was challenged with 40 μl/ml BCM for 0–24 h. (b) Immunofluorescence images showing the TLR4 localization and fluorescence distribution in BMDM, which were stimulated with BCM (40 μl/ml) for 3 and 12 h, respectively. (c) Flow cytometry showing the cell surface TLR4 expression on BMDM, which were stimulated with BCM (40 μl/ml) for 0–12 h and stained with PE-tagged anti-TLR4 antibody. (d and e) BMDM were stimulated with BCM (40 μl/ml) for 12 h followed by withdrawing BCM and continuous incubation in normal medium for 0–48 h. The cell surface TLR4 was then stained with PE-tagged anti-TLR4 antibody and detected by flow cytometry (d) and visualized by confocal microscopy using fluorescent-tagged TLR4 antibody (e). All results are representative of three independent experiments. The graph shows the mean and S.E.M., n=3. *P<0.05 and **P<0.01 versus the control