Detection of Hi-FGF-2 in the extracellular environment <i>in vitro</i> and <i>in vivo</i>.

<p><b>Panel (A)</b>. Representative western blot images of FGF-2 detection in conditioned medium from unstimulated or Ang II-stimulated hMFs. Each lane contains the heparin-sepharose-bound fraction from 60 ml of pooled conditioned medium. <b>Panel (B)</b>. Representative western blots for FGF-2 “eluted” from the cell surface with a high salt wash, and concentrated by binding to heparin-sepharose. Each lane contains the heparin-bound fraction from a 10 ml wash (5×100 near-confluent plates). Ponceau S Red (P-Red) staining of unidentified protein band(s) is also shown, indicative of equivalent loading. Experiments shown in A and B were repeated 2 more times, with similar results. <b>Panel (C)</b>. Western blot image, and corresponding quantitative data of FGF-2 isoforms present in human pericardial fluid (n = 10). Lanes 1-5 (gel 1) and 6-10 (gel 2) contain the heparin-sepharose-bound fraction from 0.5 ml pericardial fluid of individual patients. Lanes marked as F1, F2 contain recombinant Lo-FGF-2 (histidine-tagged) loaded at 0.25 and 0.5 ng/lane respectively. Sample 10 was deliberately overexposed to increase visibility of bands. Recombinant FGF-2, used as standard, was included in the second gel as well (not shown here). The graph shows percent contribution of Hi- or Lo-FGF-2 isoforms to the total FGF-2 signal (mean ± SEM). In all panels, brackets show comparisons between groups; * and ** correspond to P<0.05 and P<0.01, respectively.