Supplemental Figure S1
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Fig. S1. MT1-MMP induces LRP1 processing. (A,B) Wild-type (MT1+/+) VSMCs were cultured on polymerized collagen I for 48 hours in the presence of MMP inhibitor (GM6001, 10 µM) as indicated and treated with PDGF-BB (25 ng/ml) for 48 hours under serum-free conditions in the presence of GM6001 as above. Total cell lysates were analyzed by immunoblotting for α-chains of LRP1. (A) Whereas only the 515-kDa α-chain (also shown in Fig. 3A) was detectable with direct peroxidase conjugated anti-rabbit antibodies, (B) additional faint protein bands of <85 kDa in size became visible in the PDGF-BB-treated wild-type cells using biotin-streptavidin enhancement in the detection (indicated with arrows in B) (Lohi et al., 1996). The enhanced sensitivity also resulted in the detection of nonspecific bands of about 70, 95 and 150 kDa in size. Relative mobilities of the molecular-mass markers are indicated in kDa.
