(A) NRVMs were transduced with FAPP-PH-GFP and stimulated as indicated. Time lapse live cell microscopy was used to quantitate Golgi associated FAPP-PH-GFP fluorescence was quantitated as previously described (Zhang et al., 2013; Malik et al., 2015). NRVMs were stimulated with dobutamine alone (100 nM, left) (N = 9), dobutamine in the presence or absence of metoprolol (100 μM, center) (N = 4), or dobutamine in the presence or absence of sotalol (5 mM, right) (N = 8). Data are not significantly different between dobutamine and dobutamine + sotalol. Data are from at least 4 cells for each N. (B) NRVMs were transfected with β1-ARs and NES-Venus-mini-Gs, followed by viral transduction with CFP-Giantin. Representative confocal fluorescence images of dobutamine-mediated NES-Venus-mini-Gs recruitment (100 nM, B left), CFP-Giantin Golgi marker (B, center) and histogram of representative NES-Venus-mini-Gs recruitment (B, right). Red arrow = Perinuclear region, Black arrow = sarcolemma. The yellow line indicates where histogram data was captured. Scale bars are 10 μm. (C) Representative confocal fluorescence images of Iso-mediated NES-Venus-mini-Gs recruitment (100 nM, left), CFP-Giantin Golgi marker (B, center) and histogram of representative NES-Venus-mini-Gs recruitment. Yellow line indicates where histogram data was captured. Scale bars = 10 μm (D) Mean data of fluorescence intensity of NES-Venus-mini-Gs at the perinuclear region corresponding to the Golgi ± SEM from at least 5 cells. (E) Pearson’s correlation coefficient for overlap of YFP-Mini-Gs and CFP-Giantin images. All symbols on time course graphs are presented as mean ± standard error from N = 4 or more independent preparations of myocytes. Agonists were added where indicated by the arrow.