Figure 3.

TLR4C peptide specifically blocked the TLR4-TLR6 heterodimer in BV2 microglia cells after Aβ activation. A, A scheme showing the FRET reaction. B, representative images of cellular interaction between TLR4 and TLR6 in BV2 microglia cells with the indicated treatments observed by FRET using ImageStreamX. Scale bars, 10 μm. Cells were incubated with 20 μm concentrations of the TLR4C or scrTLR6C peptide for 0.5 h and then washed and incubated with 10 μm Aβ for another 0.5 h at 37 °C. Cells were probed with an anti-TLR6-PE-conjugated antibody (donor) and an anti-TLR4 antibody followed by staining with an APC-labeled secondary antibody (acceptor). PE intensity (middle panel) and FRET intensity (right panel) were measured. C, a graphic summary of the FRET percentage normalized to cells only. Results are the mean ± S.E. of two independent experiments (***, p < 0.005, n ≥ 17,000 for each experiment). A.U., absorbance units.