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. 2018 May 3;114(11):1499–1511. doi: 10.1093/cvr/cvy110

Figure 6.

Figure 6

The scaffold protein mAKAPβ controls β2-AR induced nuclear PKA activity when PDE4 is inhibited. (A) Immunocytochemical detection of mAKAPβ in ARVMs 72 h after sequential infection with adenoviruses encoding either a scrambled shRNA (Ad.Control shRNA MOI 2000) or a shRNA against mAKAPβ (Ad-mAKAPβ shRNA MOI 2000) for 48 h followed by infection with Ad.AKAR3-NLS (MOI 1000) for 24 h. Scale bars represent 20 µm. (B) Quantification of mAKAPβ fluorescence in ARVMs co-transduced with Ad.AKAR3-NLS and Ad.Control shRNA or Ad-mAKAPβ shRNA at 72 h. (C) Mean variation of the YFP/CFP ratio upon β1-AR stimulation with 3 nM Iso plus 10 nM ICI in ARVMs co-transduced with Ad.AKAR3-NLS and Ad.Control shRNA or Ad.AKAR3-NLS and Ad.mAKAPβ shRNA. (D) Mean variation of the YFP/CFP ratio upon β1-AR stimulation with 1 nM Iso plus 10 nM ICI in the presence of 10 µM Ro 201724 (Ro) to block PDE4. ARVMs were co-transduced with Ad.AKAR3-NLS and Ad.Control shRNA or with Ad.AKAR3-NLS and Ad.mAKAPβ shRNA. Mean variation of the YFP/CFP ratio upon β2-AR stimulation (using 30 nM Iso plus 100 nM CGP) alone (E) or in the presence of 10 µM Ro (F) in ARVMs co-transduced with Ad.AKAR3-NLS and Ad.Control shRNA or with Ad.AKAR3-NLS Ad-mAKAPβ shRNA. Number of cells/rats is indicated in brackets. Statistical significance is indicated as *P < 0.05; **P < 0.01 by nested ANOVA with Tukey’s post-hoc test.