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. 2010 Feb 22;285(17):12595–12603. doi: 10.1074/jbc.M109.008581

FIGURE 1.

FIGURE 1.

Domain structure, expression, and cleavage products of u-PAR. A, a diagram of wt u-PAR (top) shows the three domains designated D1–3 and a linker region. D3 contains the glycosylphosphatidylinositol anchor. The linker region contains the chemotactic epitope (59). A schematic of cr-u-PAR (bottom) shows the two mutated sites as underlined and in lowercase. B, immunoblotting using polyclonal rabbit α-u-PAR was used to detect total u-PAR. Samples were 125 ng of purified su-PAR as a positive control (lane 1) and lysates from non-transfected 293 cells (lane 2), 293 wt u-PAR cells (lane 3), 293 cr-u-PAR cells (lane 4), and 3-day PMA-stimulated U937 cells (lane 5). C, u-PAR cleavage products post-exposure to tcu-PA. Cells expressing wt u-PAR (lanes 1–3) or cr-u-PAR (lanes 4–6) were incubated in the absence of tcu-PA for 0 h (lanes 1 and 4) or 20 h (lanes 2 and 5) or in the presence of 100 nm tcu-PA for 20 h (lanes 3 and 6) followed by lysis, SDS-PAGE, and immunoblotting for total u-PAR.