FIGURE 4.

Analysis of LOX expression in dermal fibroblasts and skin tissue. A and B, cell lysates (A, 20 μg of total protein) and culture medium (B, 100 μl) from dermal fibroblasts of newborn littermates were separated on 4–12% polyacrylamide gels under reducing conditions and immunoblotted with a polyclonal antibody (L4669, against LOX C terminus; Sigma) recognizing both LOX precursor (pro-LOX) and mature enzyme (LOX). Short exposure of LOX in culture medium is shown (B, right panel). C, immunostaining of back skin from 4-month-old littermates with an antibody recognizing predominantly the N-terminal propeptide region and also a small segment of the mature enzyme (sc-66948, against N-terminal residues 66–205; Santa Cruz Biotechnology). Fbln4^E57K/E57K skin showed increased immunoreactivity. Tissue sections were counterstained with DAPI (blue) to visualize nuclei. Magnification bar, 50 μm. a.a., amino acids.