Extended Data Fig. 1: Golgi-bPAC stimulates cAMP generation in response to blue stimulation over time.
From: Cardiac contraction and relaxation are regulated by distinct subcellular cAMP pools
a. Representative images of HeLa cells expressing Golgi-bPAC. n = 36, 6 biological replicates. b. Concentration-response curve of cAMP generation in mouse neonatal cardiomyocytes upon various epinephrine concentrations n = 4 biological replicates. The quantified data are represented as mean ± S.E.M. c. Phosphorylation profiles of RyR2 and PLB induced by Golgi-bPAC in mouse neonatal cardiomyocytes. The protein levels of p-RyR2 Ser2808 and p-PLB Ser16/Thr17 were analyzed in the Golgi-bPAC-expressing mouse neonatal cardiomyocytes kept in the dark or exposed to 0.34 or 3.20 μW/cm2 blue light for 1.5, 3 and 7 minutes. n = 1. d. Phosphorylation profiles of RyR2, TnI and PLB induced by Golgi-bPAC before and after treatment of 20μM PKA inhibitor (H89) in mouse neonatal cardiomyocytes. The phospho protein levels were detected using of p-RyR2 Ser2808, p-TnI Ser23/24 and p-PLB Ser16/Thr17 antibodies. The protein level of Golgi-bPAC was analyzed using the SNAP antibody. The protein level of CSQ2 was used as a loading control. n = 1. e. The subcellular localization of p-PLB before blue light illumination (top row); the subcellular localization of p-PLB p-RyR2, p-TnI, upon stimulation of Golgi-bPAC with 3.20 μW/cm2 blue light in mouse neonatal cardiomyocytes (bottom rows). The protein localizations of p-RyR2 Ser2808, p-TnI Ser23/24, total PLB, and pPLB Ser16/Thr17 antibody were visualized (green) with Golgi marker stained by GM130 (red). n = 27, 3 biological replicates. Scale bar, 10 μm.
