Osteopontin mediates obesity-induced adipose tissue macrophage infiltration and insulin resistance in mice
J. Clin. Invest. Takashi Nomiyama, et al. 117:2877 doi:10.1172/JCI31986 [Go to this article.]

Figure 5
Chemotaxis of macrophages isolated from OPN^+/+ and OPN^–/– mice. (A) Peritoneal macrophages from OPN^+/+ and OPN^–/– mice were subjected to chemotaxis assays in modified Boyden chambers. Membranes of the transwell chambers were coated either with the substrate poly-D-lysine (PDL) as control or with recombinant OPN (5 ng/ml). Following attachment of the macrophages to the membrane, vehicle or MCP-1 (50 ng/ml) was added to the media in the lower chamber. Transwell migration was analyzed after 2 hours and expressed as cell numbers per HPF (×200). Experiments were repeated 4 times in triplicate. Data are expressed as mean ± SEM. *P < 0.05 compared with PDL alone; ^#P < 0.05 compared with OPN alone; ^ΧP < 0.05 compared with OPN^+/+. (B) Stromal vascular cells were isolated from epididymal adipose tissues harvested from OPN^+/+ (black bars) and OPN^–/– (white bars) mice fed a LFD or HFD for 25 weeks (n = 6/group). Cells were cultured in the bottom chambers, and peritoneal macrophages from wild-type mice were added to the insert. Migration was analyzed in triplicate after 2 hours as described in A. Data are expressed as mean ± SEM. ^†P < 0.05, compared with LFD; ^‡P < 0.05, compared with OPN^+/+ mice fed HFD.