Time dependence of RAGE shedding induced by either PACAP (A) or PMA (B).

<p>Upper figure part: representative Western blots for detection of sRAGE, full-length RAGE (RAGE fl) and actin; middle figure part: Quantitative analysis of sRAGE amounts; lower figure part: Quantification of full-length RAGE expression. PAC1/RAGE cells were incubated for different periods of time (15 min up to 360 min) using either 300 nM PACAP-27 or 1 µM PMA. sRAGE was detected and quantified as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041823#pone-0041823-g001" target="_blank">Figure 1</a>. Full-length RAGE and actin were analyzed in cell lysates by Western blotting using either the monoclonal anti-RAGE antibody 5328 or anti-actin antibody A2066. Detection was performed with ECL. For quantification of sRAGE and full-length RAGE, the experiments were done in triplicates. Shown are the mean effects ± S.D.