FIGURE 7.

Western blots of TSG-6 and mast cell tryptases in arthritic and normal paw extracts and co-immunoprecipitation of in vivo formed TSG-6-tryptase complexes. A–C, Western blots show the same membrane, which was stained with an antibody to TSG-6 (A) and then stripped and restained with antibodies to mMCP-6 (B) and mMCP-7 (C), respectively. The lanes were loaded with samples as follows: 0.2 μg of purified rmTSG-6, 0.1 μg of recombinant mMCP-6, tissue extract (50 μg of protein) from arthritic mouse paw (Arthr), extract (50 μg of protein) from normal control paw (Cntrl), and 0.05 μg of mMCP-7. D–F, extracts of arthritic (left lanes) or normal (right lanes) mouse paws were immunoprecipitated (IP) with a mixture of anti-TSG-6 mAbs (as described under “Experimental Procedures”) and then blotted (WB) with anti-TSG-6 (RC21) (D), anti-mMCP-6 (E), and anti-mMCP-7 antibodies (F). The arrows point to the highest and lowest molecular weight TSG-6 species in D. The broken arrow indicates the TSG-6 coimmunoprecipitated mMCPs.