Article Figures & Data
Additional Files
- Supplemental Data -
Supplemental Figure 1: Endothelium-independent effect of cinaciguat
Intact and endothelium-denuded rings from porcine coronary arteries were incubated
with 1 nM (A), 10 nM (B), or 100 nM (C) cinaciguat and relaxation was recorded up to
90 min. Vascular response to 1 μM bradykinin was measured to test for the
effectiveneß of endothelial removal (D). Data represent mean values ± S.D. of
coronary rings obtained from 3 individual hearts; *p<0.05 (unpaired t-test).Supplemental Figure 2: Effect of cinaciguat on vascular PDEs
Homogenates prepared from porcine coronary arteries were preincubated for 10, 40,
and 90 min in the absence and presence of cinaciguat (1 μM) with or without IBMX (1
mM) or the PDE5-specific inhibitor sildenafil (1 μM; data not shown) at 37 °C. At the
indicated time points, 10 μM [32P]cGMP was added to the reaction mixture and
incubated for 10 min. PDE activity was determined as hydrolysis of [32P]cGMP. Data
represent mean values ± S.D. of 3 experiments performed in duplicate; n.d. not
detectable.Supplemental Figure 3: Effect of cinaciguat on cGMP formation in cells
Porcine endothelial cells were incubated for 45 min under basal conditions and in the
presence of 10 μM or 100 μM cinaciguat in serum-free DMEM. Thereafter, reactions
were either terminated by removal of medium and addition of 10 mM HCl or, for
washout experiments, medium was replaced by fresh DMEM and cells were
incubated for further 60 min. Data represent mean values ± S.D. of 4 independent experiments performed in triplicate. DEA/NO (1 μM) induced formation of 52,36 ± 39,07 pmol cGMP per 106 cells under control conditions.Supplemental References
Schmidt K, Mayer B and Kukovetz WR (1989) Effect of calcium on endotheliumderived
relaxing factor formation and cGMP levels in endothelial cells. Eur J
Pharmacol 170(3):157-166.Schultz G and Böhme E (1984) Guanylate Cyclase GTP pyrophosphate-lyase
(cyclizing), in Meth Enzym Anal (Bergmeyer HU, Bergmeyer J and Graßl M eds) pp
379 - 389, Verlag Chemie, Weinheim.
- Supplemental Data -