FIG. 5.

LOX enzyme activity within pooled media (A) or within cell matrix (B), when GSNO (0–100 nM) was supplemented to HA-SMC cultures, in the presence or absence of HA-EC cocultures. LOX activity was quantified using a fluorometric AmplexRed^® assay. Data shown represent mean±standard error of LOX synthesis after 21 days of culture, normalized to cell count within respective cases (n=6 per condition). Data from western blot analysis showed fold increases in expression of elastin protein (C) and LOX protein (D) within pooled media, compared with GSNO-free cultures (controls). Representative SDS-PAGE/Western blots containing bands corresponding to tropoelastin and LOX protein in pooled media are shown for comparison. The protein bands were quantified in respective cases (n=3 per condition) and normalized to controls. *p<0.05 compared with GSNO-free cultures; ^#p<0.01 for no cocultures versus EC coculture, at a given GSNO dosage.