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. 2005 Apr 5;387(Pt 2):497–506. doi: 10.1042/BJ20041324

Figure 2. MT1-MMP forms shed by surface biotinylated HT1080 cells.

Figure 2

(A) Serum-free conditioned media from PMA (‘TPA’)- (lane 1) or Con A-treated (lane 2) HT1080 cells were incubated with the MMP inhibitor-tethered matrix and the bound proteins were detected by immunoblot (IB) analysis using mAbLEM-2/15. (B) Untreated (lanes 1, 3, 5 and 7) or Con A-treated (lanes 2, 4, 6 and 8) HT1080 cells were surface biotinylated (lanes 3, 4, 7 and 8) or non-biotinylated (lanes 1, 2, 5 and 6). The serum-free conditioned media were collected, incubated with the inhibitor-tethered matrix and the bound fractions were subjected to SDS/PAGE (12% reducing gel) and transferred on to a nitrocellulose membrane. The same membrane was developed with streptavidin/HRP (lanes 1–4) and with mAbLEM-2/15 (lanes 5–8). (C) Serum-free media of MT1-MMP expressing BS-C-1 cells were incubated with the inhibitor-tethered matrix in the absence or presence of recombinant TIMP-2 or TIMP-1. The 50 kDa form of MT1-MMP in the unbound and bound fractions were detected by immunoblot analysis using the mAbLEM-2/15.