Phospholipase C β3 deficiency leads to macrophage hypersensitivity to apoptotic induction and reduction of atherosclerosis in mice
J. Clin. Invest. Zhenglong Wang, et al. 118:195 doi:10.1172/JCI33139 [Go to this article.]

Figure 1
Effects of PLC β3 deficiency on macrophage functions. (A) Calcium effluxes induced by 10 nM C5a in macrophages isolated from wild-type mice and mice lacking PLC β2 (P₂), PLC β3 (P₃), or both (P₂₃). (B–E) Transendothelial migration of peritoneal macrophages and spleen monocytes. Peritoneal macrophages (B and C) or splenocytes (D and E) were added to the top chambers of Transwell plates precoated with mouse endothelial cells. C5a (10 nM), SDF-1 (100 ng/ml), or MCP-1 (60 ng/ml) were added to the lower chambers. After 4-hour incubation, cells in the lower chambers were collected, counted, and analyzed by FACS after staining with F4/80 (B and C) or CD11b/Gr1 (D and E) antibodies. The indices were calculated by dividing the number of F4/80-positive (B and C) or Gr-1^loCD11b^mid (D and E) cells in the presence of ligand by that in its absence. n = 8; P > 0.05 for all B–E. (F and G) Adhesion of spleen monocytes to extracellular matrices and endothelial cells. Spleen cells were added onto 24-well plates without coating or coated with poly-lysine, fibronectin (F), or mouse endothelial cells (G). After 10 minutes, attached cells were collected, counted, and analyzed by FACS for Gr-1^loCD11b^mid monocytes. n = 4; P > 0.05 for F and G. (H) Macrophage phagocytosis. Peritoneal macrophages were seeded on 24-well culture plates. Uncoated and oxLDL-coated FluoSphere beads were added to the cells. After extensive washes, cells were detached, stained with F4/80, and analyzed by FACS. n = 4; P > 0.05 between genotypes. (I) Uptake of apoptotic Jurkat cells. Macrophages were incubated with CFDA SE–labeled apoptotic Jurkat cells, detached, stained with a macrophage marker F4/80, and analyzed by FACS. The MFI of CFDA SE associated with F4/80-positive cells was determined. n = 4; P > 0.05. fn, fibronectin.