Effects of IGF-1 stimulation on the interaction between NCS-1 and IP₃Rs in cardiomyocytes.

<p>(A and B) Interactions between NCS-1 and IP₃Rs were visualized and quantified using <i>in situ</i> proximity ligation assay (PLA) in α-actinin-positive NMVMs expressing NCS-1-HA. (A) Representative <i>in situ</i> PLA images of NCS-1-HA/IP₃R interactions (detected as white dots) before and 5 or 10 min after IGF-1 (10 nmol/L) treatment. α-Actinin-positive cells (myocytes) were stained green, and nuclei were stained blue by DAPI. White arrowheads indicate the signals with close proximity to the nuclear membrane. Experiments were also carried out without primary antibodies as a negative control (leftmost). Bar, 10 μm. (B) Quantification of the NCS-1-HA/IP₃R interactions. The number of interactions/cell was plotted for all the cells and the mean values for each group are presented as bars. *<i>P</i> < 0.05, **<i>P</i> < 0.01 vs. IGF-1 0 min; n = 80–88 in three independent experiments.