1. Clustering of CD14 in the plane of the plasma membrane of LPS-stimulated cells.
Clustering of CD14 in the plane of the plasma membrane of LPS-stimulated cells. J774 cells were left unstimulated (A,E) or exposed to 100 ng/ml LPS for 5 min (B,F), 30 min (C,G) or 60 min (D). Subsequently, they were incubated with a rat antibody against CD14 followed by goat anti-rat IgG conjugated to 10-nm gold (A–D), or CTX followed by a rabbit antibody against CTX and goat anti-rabbit IgG conjugated to 6-nm gold (E–G) to visualize CD14 and GM1, respectively. After surface labeling, plasma membrane sheets were prepared through mechanical cleavage of cells and examined under an electron microscope. In A–D, CD14 singlets and doublets, and clusters are marked with small and large circles, respectively. Scale bar: 500 nm. (H) Distribution of cluster sizes of gold particles marking CD14 and GM1 before and during stimulation of cells with 100 ng/ml LPS. Data are expressed as the percentage of total gold particles found at a given time point on a 100-μm2 area of the plasma membrane. The mean±s.e.m. of three or four experiments is shown.
Agnieszka Płóciennikowska et al. J Cell Sci 2015;128:
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