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. Author manuscript; available in PMC: 2022 Jan 22.
Published in final edited form as: Circ Res. 2020 Nov 13;128(2):246–261. doi: 10.1161/CIRCRESAHA.120.317452

Figure 2. Differential local regulation of NE-induced β1AR/PKA activities in WT and OCT3KO AVMs.

Figure 2.

(A-B) Schematics of local activation of β1AR-induced PKA activity at subcellular locations and detection using FRET based biosensors (plasma membrane, PM-AKAR3 and sarcoplasmic reticulum, SR-AKAR3) in WT and OCT3KO AVMs. Isoproterenol (ISO), norepinephrine (NE). FRET assay was analyzed with F/F0 of YFP/CFP ratio. (C-D) Concentration-response curves of changes in YFP/CFP ratio after ISO stimulation in WT and OCT3KO AVMs expressing PM-AKAR3 or SR-AKAR3. Data were from 6 WT and 6 OCT3KO mice. (E-F) Concentration-response curves of changes in YFP/CFP ratio after NE stimulation in WT and OCT3KO AVMs expressing PM-AKAR3 or SR-AKAR3. Data were from 8 WT and 7 OCT3KO mice. Data are shown in mean ± S.E.M. p values were obtained by two-way ANOVA with Tukey’s multiple comparison test when comparing WT to OCT3-KO. (G-H) Detection of phosphorylation of phospholamban (PLB) at serine 16 and phosphorylation of phospholemman (PLM) at serine 63 in mouse AVMs after stimulation with 100 nmol/L ISO or 100 nmol/L NE. Data are shown in mean ± S.E.M. (N = 5). A.U. (arbitrary unit) is defined as the ratio of intensity of phosphorylated proteins over intensity of total proteins. p values were obtained by two-way ANOVA with Tukey’s multiple comparison test.