L name

Losartan, PD123319 and Angiotensin II were purchased from Sigma-Aldrich Co. (St. Louis, USA). L-NAME was purchased from Santa Cruz Biotechnology (Dallas, USA). C21 was provided by Vicore Pharma AB (Göteborg, Sweden). Doses were selected based on previous studies [14 (link),25 (link),26 (link)].

TPPU was purchased from Cynthia labs (Davis, CA, USA), and L-NAME from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The reagents and kits for Western Blot were purchased from Abcam (Burlingame, CA, USA). Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and membranes were purchased from Bio-Rad (Des Plaines, IL, USA). PCR kits and reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Vasoactive signaling pathways were inhibited mice with the following protocols that have been previously validated. Inhibition of eNOS was performed with 75 μg/kg L-nitroarginine methyl ester (L-NAME, Santa Cruz Biotech., Santa Cruz, CA) given IP 30 min prior to study. Blockade of adenosine A₂-receptors was performed with 50 μg/g of 4-(2-[7-Amino-2-(2-furyl)[1,2,4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl)phenol (ZM-241385, Abcam PLC, Cambridge, MA) given IV 1 hr prior to study. This dose is predicted to inhibit both A_2a and A_2b receptors. Accelerated metabolism of ATP was performed with 4.0 U/g apyrase (Sigma-Aldrich, St. Louis, MO) given IP 1 hour prior to study; while inhibition of ectonucleotidases involved in ATP metabolism was performed by 1 μg/g of sodium polytungstate (POM-1, VWR, Radnor, PA) given by IP injection 30 min prior to study. For inhibition of the K_ATP receptor was performed by 15 mg/kg glibenclamide (Santa Cruz Biotechnology, Dallas, TX) given IV 30 min prior to study. For inhibiting intracellular signaling pathways, L-NAME (20 mg/kg) was administered by IV route 10 min prior to imaging; and indomethacin (2 mg/kg) was given by IP route 24 hrs and again 1 hr prior to study.