Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain (Mus musculus), C57BL/6J |
wild-type | Jackson Laboratories | stock # 000664 | |
| Strain (Mus musculus) |
S1928A | (Lemke et al., 2008) | ||
| Cell line (human embryonal kidney) |
tsA-201 | Sigma-Aldrich | 96121229 | SV40 transformed |
| Oligodeoxynucleotide, sense | P2Y11 SNS ODN | Integrated DNA Technologies | human P2Y11 sequence (NC_000019.10): 5’-CAACGTCTCGGGTAAGGAGAA-3’ and 5’-ATGAGGAAGGAAACGTGGGT-3’ |
last three bases on the 3’ end were phosphoro-thioated |
| Oligodeoxynucleotide, antisense | P2Y11 ANS ODN | Integrated DNA Technologies | human P2Y11 sequence (NC_000019.10): 5’-CAAGGCCACCCTAACCACTG-3’ and 5’-CTCTCCCTTCCCTGCGTTA-3’ |
last three bases on the 3’ end were phosphoro-thioated |
| DNA construct | human P2Y11 | UMR cDNA Resource Center |
www.cDNA.org; clone ID P2Y1100000 |
tagged with GFP at C- terminus |
| Antibody | anti-FP1 (CaV1.2; custom rabbit) | (Davare et al., 2000) | dilutions: 1:100 for immunoblot and PLA; 10 µg/mL for GSD |
|
| Antibody | anti-CH3P (pSer1928, custom rabbit) |
(Davare et al., 2000) | 1:50-1:100 dilutions | |
| Antibody | anti-β-actin (mouse monoclonal) | Abcam | ab8226 RRID: AB_30637 |
1:1000 dilution |
| Antibody | anti-α-tubulin (mouse monoclonal) | Active Motif | 39527; clone 5-B-1–2 | 1:500 dilution |
| Antibody | anti-P2Y11 (rabbit polyclonal) | Abcam | ab180739 | 1:100-1:200 dilutions |
| Antibody | anti-P2Y11 (goat polyclonal) | Santa Cruz Biotechnology |
sc-69588; clone C-18 RRID: AB_21559 |
dilutions: 1:100 for stainings (PLA, classical) and 10 µg/mL for GSD |
| Antibody | anti-PKAcatα, β, γ (mouse monoclonal) |
Santa Cruz Biotechnology |
sc-28315; clone A-2 | 1:200 dilution |
| Antibody | PKAcatα, β, γ blocking peptide | Santa Cruz Biotechnology |
sc-28315 P; clone A-2 | 1:20 dilution for 1 µg of primary antibody |
| Antibody | anti-PKAcat α, β, γ (rabbit polyclonal) | Santa Cruz Biotechnology | sc-28892; clone H-95 | dilutions: 1:200 for PLA and 10 µg/mL for GSD |
| Antibody | Alexa Fluor 488 conjugate of wheat germ agglutinin | Life Technologies | W7024 | |
| Antibody | Alexa Fluor 568-conjugated donkey anti-goat | Molecular Probes | A11057 RRID: AB_142581 |
5 mg/mL dilution |
| Antibody | Alexa Fluor 568-conjugated donkey anti-mouse | Molecular Probes | A10037 | 5 mg/mL dilution |
| Antibody | Alexa Fluor 568-conjugated donkey anti-rabbit | Molecular Probes | A11011 RRID: AB_143157 |
2 µg/mL dilution |
| Antibody | Alexa Fluor 647-conjugated donkey anti-goat | Molecular Probes | A21447 RRID: AB_141844 |
2 µg/mL dilution |
| Antibody | goat anti-rabbit IgG (H + L)-horseradish peroxidase conjugate | Bio-Rad | 170–6515 RRID: AB_11125142 |
1:10000 dilution |
| Antibody | goat anti-mouse IgG (H + L)-horseradish peroxidase conjugate | Bio-Rad | 170–6516 | 1:10000 dilution |
| Chemical compound, drug |
sodium pentobarbital (Fatal-Plus) | Vortech Pharma-ceuticals | NDC 0298-9373-68 | |
| Chemical compound, drug |
mannitol | Fisher Scientific | BP686 | |
| Chemical compound, drug |
NF340 | Santa Cruz Biotechnology | sc-361274 | |
| Chemical compound, drug |
NF546 | Tocris | 3892 | |
| Chemical compound, drug |
apyrase | New England Biolabs |
M0398L | |
| Chemical compound, drug |
nifedipine | Sigma-Aldrich | N7634 | |
| Chemical compound, drug |
Bay K-8644 | Sigma-Aldrich | 71145-03-4 | |
| Chemical compound, drug |
forskolin | Sigma-Aldrich | F6886 | |
| Chemical compound, drug |
amphotericin B | Sigma-Aldrich | A4888 | |
| Chemical compound, drug |
MRS2578 | Santa Cruz Biotechnology | sc-204103A | |
| Chemical compound, drug |
MRS2179 | Tocris | 1454889-37-2 | |
| Chemical compound, drug |
protein kinase A inhbitor (PKI) | Sigma-Aldrich | P9115 | fragment 14–22, myristoylated trifluoroacetate salt |
| Chemical compound, drug |
Rp-Adenosine 3’,5’-cyclic monophos-phorothioate triethylam-monium salt (rpcAMP) |
Sigma-Aldrich | A165 | |
| Chemical compound, drug |
Adenosine 5’-[γ-thio] triphosphate tetralithium salt (ATPγS) | Sigma-Aldrich | A1388 | |
| Software, algorithm |
GraphPad Prism | GraphPad Prism, RRID: SCR_002798 | ||
| Software, algorithm |
ImageJ | Fiji, RRID: SCR_002285 |
||
| Software, algorithm |
pCLAMP10 | Molecular Devices | electrophysiology | |
| Software, algorithm |
LASAF | Leica | GSD | |
| Software, algorithm |
IonOptix | IonOptix | arterial diameter recordings |
|
| Software, algorithm |
Metaflor | Molecular Devices | FRET |