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. 2022 Aug 1;11:e77725. doi: 10.7554/eLife.77725

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© 2022, Shen et al

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Figure 2. — (a–c) Representative Ca²⁺ release units (CRUs) imaged in cardiomyocytes treated with isoproterenol (100 nM), 8-CPT (10 µM), or caffeine (0.5 mM) for 1 hr (left panels) or with inclusion of the CaMKII inhibitor AIP (2 µM) for an additional hour (right panels). Scale bar: 100 nm. (d– f) Induced RyR cluster dispersion was reversed by CaMKII inhibition, as indicated by measurements of RyR cluster size, CRU size, and cluster density (control: n_cells = 50, n_hearts = 6; Iso: n_cells = 37, n_hearts = 5; Iso + AIP: n_cells = 37, n_hearts = 5; 8-CPT: n_cells = 48, n_hearts = 5; 8-CPT + AIP: n_cells = 52, n_hearts = 5; caffeine: n_cells = 42, n_hearts = 4; caffeine + AIP: n_cells = 35, n_hearts = 3). (g–i) Representative images of RyR organization in cardiomyocytes from wildtype (WT) mice and transgenic mice with constitutively activated (S2814D) or genetically ablated (S2814A) phosphorylation at S2814. Clusters were cropped from a region measuring 5 x 9 x 0.6 μm. Images and quantified experimental data (j–l) are presented under baseline conditions and following 60 min isoproterenol stimulation (WT: n_cells = 25, n_hearts = 2; WT + Iso: n_cells = 24, n_hearts = 2; S2814D: n_cells = 19, n_hearts = 2; S2814D + Iso: n_cells = 18, n_hearts = 2; S2814A: n_cells = 17, n_hearts = 2; S2814A + Iso: n_cells = 19, n_hearts = 2). The bar charts present mean measurements ± SEM, with superimposed data points representing averaged values from each cardiomyocyte. Statistical significance (p<0.05) between groups is indicated by a comparison bar.

Figure 2—figure supplement 1. — (a–c) Representative Ca²⁺ release units (CRUs) imaged in cardiomyocytes treated with isoproterenol (100 nM), 8-CPT (10 µM), or caffeine (0.5 mM) for 1 hr (left panels) or with inclusion of the CaMKII inhibitor AIP (2 µM) for an additional hour (right panels). Scale bar: 100 nm. (d– f) Induced RyR cluster dispersion was reversed by CaMKII inhibition, as indicated by measurements of RyR cluster size, CRU size, and cluster density (control: n_cells = 50, n_hearts = 6; Iso: n_cells = 37, n_hearts = 5; Iso + AIP: n_cells = 37, n_hearts = 5; 8-CPT: n_cells = 48, n_hearts = 5; 8-CPT + AIP: n_cells = 52, n_hearts = 5; caffeine: n_cells = 42, n_hearts = 4; caffeine + AIP: n_cells = 35, n_hearts = 3). (g–i) Representative images of RyR organization in cardiomyocytes from wildtype (WT) mice and transgenic mice with constitutively activated (S2814D) or genetically ablated (S2814A) phosphorylation at S2814. Clusters were cropped from a region measuring 5 x 9 x 0.6 μm. Images and quantified experimental data (j–l) are presented under baseline conditions and following 60 min isoproterenol stimulation (WT: n_cells = 25, n_hearts = 2; WT + Iso: n_cells = 24, n_hearts = 2; S2814D: n_cells = 19, n_hearts = 2; S2814D + Iso: n_cells = 18, n_hearts = 2; S2814A: n_cells = 17, n_hearts = 2; S2814A + Iso: n_cells = 19, n_hearts = 2). The bar charts present mean measurements ± SEM, with superimposed data points representing averaged values from each cardiomyocyte. Statistical significance (p<0.05) between groups is indicated by a comparison bar.