Effect of Ca²⁺ on the shedding of RAGE.

<p>PAC1/RAGE cells were pre-incubated for 1 h with respective inhibitor and then 300 nM PACAP-27 was added for 2 h. Secreted RAGE in the cell culture supernatant 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>. Shown are the mean effects ± S.D., significance was determined by the One-way ANOVA test (ns = P>0.05; ** = P<0.01; *** = P<0.001). (A) Effect of protein kinase C inhibitors Gö6976 and Gö6983 (both 1 µM); (B) effect of the IP3 receptor modulator 2-APB (75 µM), and (C) effect of the calcium/calmodulin-dependent protein kinase II inhibitor CK59 (20 µM). In all experiments effects of inhibitors were also analyzed in the absence of PACAP stimulation.