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. Author manuscript; available in PMC: 2020 May 13.
Published in final edited form as: Sci Transl Med. 2019 Jul 10;11(500):eaav5464. doi: 10.1126/scitranslmed.aav5464

Fig. 2: A-71915 is a partial agonist of mNPR1.

Fig. 2:

(A) Schematic depicting our strategy to measure NPR1 activity. NPR1 is stimulated by NP to increase synthesis of cGMP, in turn, increased cGMP alters the conformation of a PDE5-Firefly luciferase-based sensor (cGMP-sensor) which results in hydrolysis of GloSensor reagent and production of light. (B) Time course experiments quantifying luminescence of HEK-293 cells transiently expressing cGMP-sensor stimulated with the soluble GC activator SNP (333 μM; red), media (black), hNPPA (blue), hNPPB (green), and NPPC (purple)(10 nM each). (C) Quantification of activity of HEK-293 cells transiently expressing mNPR1 and cGMP-sensor stimulated with mNPPB (green), mNPPA (blue) and NPPC (purple). (D) Quantification of inhibition of mNPR1-cGMP-sensor cells with A-71915 (5 minutes after addition of A-71915, cells were treated with 1 nM mNPPB). (E) Quantification of mNPR1-cGMP-sensor cells shows partial agonist activity for A-71915. Data represent means ±  SEM of triplicate (B) or duplicate (C-E) measurements.