Fig 5.
CD36-ligand induced TLR4-TLR6 complex formation. (a-b) Immunoblot analysis of CD36-precipitated proteins from THP-1 monocytes treated with oxLDL (50 μg/ml) or Aβ1-42 (50 μg/ml) for the indicated times. Precipitated proteins were immunoblotted with antibodies to TLR4, TLR6 and CD36. (c) Fluorescence microscopy of HEK293 cells transfected with CD36 and fluorescently tagged TLR4-YFP and TLR6-CFP. Cells were unstimulated (left) or treated with oxLDL (right) for 30 min, and stained with an anti-CD36 Alexa647 conjugated antibody. In the false-colored merged image triple co-localization of TLR4 (blue), TLR6 (red) and CD36 (green) appears as areas of white (arrowheads). Expression of individual receptors is shown in bottom panels. Scale bar 10 μm. (d-e) Inhibition of dynamin-dependent endocytosis blocks CD36-TLR4-TLR6-signaling in macrophages and microglia. QRT-PCR analysis of gene expression in (d) macrophages stimulated with oxLDL (50 μg/ml, 6h) or (e) microglia stimulated with Aβ1-42 (10 μM, 6h) in the presence and absence of the dynamin inhibitor dynasore (80 μM).