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. Author manuscript; available in PMC: 2009 Feb 25.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2007 Apr 5;27(6):1447–1455. doi: 10.1161/ATVBAHA.107.143438

Figure 1.

Figure 1

Localization of tissue factor and PAR2 in lipid rafts. A, Presence of TF and PAR2 antigens in low-density, detergent-insoluble microdomains. MDA-MB-231 cells were lysed with 1% Triton X-100 and fractionated on sucrose gradient centrifugation. Equal amounts of protein from each fraction were separated on SDS-PAGE and transferred for immunoblotting with polyclonal antibodies to TF and PAR2, mAB to caveolin-1, and HRP-conjugated cholera toxin. Fractions are labeled from the top to the bottom of the sucrose gradient. B, Immunofluorescence microscopy of TF, PAR2, and caveolin-1. MDA-MB-231 cells were fixed, and intact (for TF and PAR2 colocalization) or permeabilized cells were stained with polyclonal or monoclonal antibodies to TF, PAR2, and caveolin-1. Right panels depict overlay of images, and yellow in the panel represents colocalization of TF and PAR2 with caveolin-1. The images shown are montages of z-stacks.