Runcaciguat

The drugs used in this experiment—acetylcholine (ACh), phenylephrine (PE), L-NAME (NΜ-nitro-L-arginine methylester hydrochloride)—were purchased from Sigma, St. Louis, MO, USA; SNP was purchased from Carl Roth, Karlsruhe, Germany; sildenafil citrate from BioVision, Milpitas, CA, USA. Runcaciguat was provided by Bayer AG, Research & Development, Pharmaceuticals (Wuppertal, Germany). All salts, glucose and HEPES used to prepare buffer solutions were purchased from Carl Roth, Karlsruhe, Germany. PE and ACh were dissolved in distilled water and stored at −20 °C. The SNP solution was prepared in distilled water immediately before use and stored in an amber tube. L-NAME was dissolved using ultrasound in distilled water. Runcaciguat was initially prepared in dimethylsulphoxide (DMSO, Thermo Scientific, Bellefonte, PA, USA) and stored at −20 °C.

BAY-747 and runcaciguat were supplied by Bayer AG (Leverkusen, Germany)^{25 (link),26 (link)}. Donepezil, an acetylcholinesterase inhibitor (AChEI), was generously donated by Abbott (Weesp, The Netherlands). Methyl 2-hydroxyethyl cellulose (Tylose^® MH300) and Tween80 (polyoxyethylenesorbitan monooleate, cat# P8074) were purchased from Sigma-Aldrich Chemie bv (Steinheim, Germany). Forskolin was purchased from Tocris Bioscience (#1099, Abingdon, UK), rolipram was purchased from Abcam (#ab120029), and sulfo-NHS-SS-biotin (#A39258) and streptavidin-coated Dynabeads (#65601) were purchased from Thermo Scientific (Bleiswijk, The Netherlands).
Mouse anti-GluA1 was purchased from Merck Millipore (#MAB2263, Burlington, MA, USA). Rabbit anti-GluA1 phospho S845 was purchased from Abcam (#ab76321), and mouse anti-GAPDH was purchased from Fitzgerald Industries (#10R-G109A, Acton, MA, US) (all three antibodies successfully used previously^{13 (link),20 (link)}). Rabbit anti-TrkB (#4603S, Cell Signaling Technologies) and mouse anti-PSD95 (#56452, QED Bioscience) were a generous gift from Bayer (as used previously^{27 (link),28 (link)}).

The sGC activator runcaciguat (BAY 1101042), was synthetized at Bayer AG, Wuppertal, Germany (Hahn et al. 2021 (link)). runcaciguat was either dissolved in vehicle (transcutol, 20% cremophor, and 70% water (Ren2TG study and Ang-SD study)) and prepared freshly before administration or was mixed in the food (ZDF study).