St ht31p

mpkCCD_cl4 cells (gift from Alain Vandewalle, Paris) were cultured as previously described^{25 (link)}. The mpkCCD cells were seeded on semipermeable filters (Transwell, 0.4-µm pore size; Corning Costar). For immunofluorescence and western blot analysis, 0.33-cm² and 4.67-cm² filters were used, respectively. The mpkCCD cells were cultured for 5 days, with changes of the medium daily. St-Ht31 (Promega Corporation, Madison, WI, USA), St-Ht31P (Promega Corporation), FMP-API-1 (Sigma–Aldrich Corporation, St. Louis, MO, USA), FMP-API-1/27, FMP-API-1/28, dDAVP (Sigma–Aldrich Corporation), and H89 (Sigma–Aldrich Corporation) were applied to the basolateral side of the mpkCCD cells.

H89, NKH 477, 8-Br-cAMP, and 8-Br-cGMP were from R&D Systems (Abingdon, Oxford, UK). Sp-cAMPS, 6-Bnz-cAMP, 8-pCPT-2′-O-Me-cAMP, Rp-cAMPS, Rp-8-CPT-cAMPS, ESI-09, and HJC0197 were from Biolog (Bremen, Germany). Ionomycin, SQ 22536, DDA and myristoylated-PKA inhibitor peptide (PKI) were from Merck-Millipore (Watford, UK). A membrane-permeant peptide inhibitor of A kinase-anchoring proteins (AKAPs) [stearated Ht31 AKAP inhibitor peptide (st-Ht31)] and its proline-modified inactive form (st-Ht31P) were from Promega (Southampton, UK). Thapsigargin was from Alomone Laboratories (Jerusalem, Israel). PAR1 peptide, histamine dihydrochloride, forskolin, IBMX, and PGE₂ were from Sigma-Aldrich (Welwyn Garden City, UK). Butaprost (free acid) and L902,688 were from Cayman Chemicals (Ann Arbor, MI). Membrane-permeant caged IP₃ (ci-IP₃PM) was from SiChem (Bremen, Germany). [2,8-³H] adenine ci-IP₃PM, d-2,3-O-isopropylidene-6-O-(2-nitro-4,5-dimethoxy)benzyl-myo-inositol 1,4,5-trisphosphate-hexakis(propionoxymethyl) ester was from Perkin Elmer (Seer Green, Bucks, UK). Fluo-8 was from Stratech Scientific Ltd (Newmarket, Suffolk, UK). Other reagents were from Sigma-Aldrich, sources specified previously (Pantazaka et al., 2013 (link)) or identified in this section.

φ450-cAMP, φ575-cAMP, φ575-OMe-cAMP and φ450-free dye were either purchased or obtained as a gift from Biolog Life Science Institute (Bremen, Germany); MitoTracker Red was purchased from Life Technologies (Carlsbad, CA); EGFP from Biovision Inc. (Milpitas, CA), ST-Ht31 and ST-Ht31P from Promega (Madison, WI). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).