Characterization of the PLN p.Arg14del Mutation in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Figures

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  Figure 1. Parameters of V-induced CaT and SR Ca2+-content in WT and MUT. (a) CaT amplitude (WT n = 37, MUT n = 32); (b) CaT tpeak (WT n = 37, MUT n = 32); (c) CaT decay t1/2 (WT n = 43, MUT n = 36); (d) amplitude of caffeine-induced Ca2+ transients, reflective of SR Ca2+-content (WT n = 20, MUT n = 27); Data are expressed as mean ± SD; * p < 0.05 and **** p < 0.0001 vs. MUT.

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  Figure 2. Distribution of CaT profiles and thapsigargin effect in WT and MUT. (a) CaT profiles from the WT and MUT groups are pooled and plotted according to their decay t1/2 vs. rise time. Data points are colored according to previous (blinded) profile assignement to a given type (T1, purple; T2, black; T3, green; T4, blue; T5, red). (b) Distribution of CaT profile types in WT (left) and MUT (right) preparations; profiles representative of each type are shown below the axis; (c) Representative WT (left) and MUT (right) CaT profiles in basal condition (TYR, black line) and after thapsigargin (THAPSI, red line).

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  Figure 3. PST3093 1µM (PST) effect on V-induced CaT parameters and SR Ca2+-content in WT and MUT. (a) CaT amplitude in WT (TYR n = 37, PST n = 13) and MUT (TYR n = 32, PST n = 28); (b) tpeak in WT (TYR n = 37, PST n = 25) and MUT (TYR n = 32, PST n = 28); (c) Decay t1/2 in WT (TYR n = 43, PST n = 34) and MUT (TYR n = 36, PST n = 28); (d) SR Ca2+-content in WT (TYR n = 20, PST n = 12) and MUT (TYR n = 27, PST n = 28). Data are expressed as mean ± SD; * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. TYR.

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  Figure 4. PST3093 1µM (PST) effect on the distribution of CaT profiles. Effect of PST on CaT profiles in WT (panel (a); TYR left, PST right) and MUT (panel (b); TYR left, PST right).

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  Figure 5. Rate-dependency of CaT parameters and CaD. Rate-dependency was tested by stepwise increments in pacing rate (1 Hz to 1.3 Hz, 1.7 Hz, and 2 Hz), the last 3 beats at steady-state are shown for each rate. (a) WT (black) vs. MUT (grey) in basal conditions (TYR; 9 ≤ n ≤ 15 for both groups); (b) PST3093 1µM (PST) effect in WT (TYR black, PST purple, 9 ≤ n ≤ 20); (c) PST effect in MUT (TYR grey, PST cyan; 9 ≤ n ≤ 20. Data are expressed as mean ± SD; * p < 0.05 for the interaction factor at two-way ANOVA.

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  Figure 6. Rate-dependency of CaT profile in WT and MUT. Representative CaT profiles for WT and MUT in basal conditions and in presence of PST3093 1 µM (PST) at pacing rates of 1 Hz (a), 1.3 Hz (b), 1.7 Hz (c), and 2 Hz (d). The rising phase of CaT is shown at a faster time scale in the inset of each panel.

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  Figure 7. Subcellular PLN and SERCA2a localization in WT and MUT. Representative PLN and SERCA2a immunolabeling images (left) and the respective intensity profiles (distance from nucleus center expressed as % of total) for short and long cell axis (right) are shown. In the intensity profiles, the horizontal bars identify the perinuclear (N) and cytosolic (C) regions. (a) WT, (b) MUT. S = short axis, L = long axis.

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  Figure 8. Average PLN and SERCA2a subcellular intensity profiles in WT and MUT. The PLN and SERCA2a intensity profiles (in the short and long cell axis) were binned and averaged among cells. WT (black, n = 9) and MUT (grey, n = 9). Subcellular regions are identified by the dotted lines: PeriNuc = perinuclear; Cyt = cytosolic; SubMem = subsarcolemma; Mem = sarcolemma. * p < 0.05 vs. MUT (by two-way ANOVA).