SUPPLEMENT 2This power point presentation reports analyses of RyR distribution
measured by confocal microscopy in tangential sections of 6 SA node cells
Summary of the analyses
* Cell numbers are those assigned for 25 cells reported in Supplemental Figure 1
*Cell#, section All peaks Peaks with higher amplitudes (> Mean+2 x SD)**
Number of peaks
#peaks withover-micronneighbors
Nearest neighbor
distance, mmMean±SD
Number of peaks
#peaks withover-micronneighbors
Nearest neighbor
distance, mmMean±SD
Cell#5,bottom
319 79 (24.8%) 0.839±0.225 20 20 (51.3%) 1.186±1.130
Cell#10,bottom
116 7 (6.0%) 0.710±0.223 31 18 (58.1%) 1.044±0.474
Cell#8,top
122 35 (28.7%) 0.871±0.273 18 11 (61.1%) 1.863±1.188
Cell#11,top &bottomcombined
365 118 (32.4%) 0.888±0.300 74 43 (58.1%) 1.255±0.831
Cell#13,bottom
153 32 (20.9%) 0.775±0.276 47 27 (57.5%) 1.112±0.529
Cell#19,bottom
149 21 (14.1%) 0.719±0.289 39 20 (51.3%) 1.186±1.130
Average ± SEM
(21.2±4.0%) 0.800±0.03 (56.2±1.6%) 1.274±0.121
** The SD in finding peaks with higher amplitude (>mean+2 x SD) was calculated for fluorescence peaks given in the units of the SD of the total fluorescence signal
original
cell#5_bottom
20 mm
5*
cell#5_bottomIntensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
cell#5_bottom
Results
All peaks
Nearest neighbors distancemean= 1.186087SD= 1.1298
# peaks total 39# >1 mkm neighbor 20% >1 mkm neighbor 51.28205
Nearest neighbors distancemean= 0.8389SD= 0.224932
# peaks total 319# >1 mkm neighbor 79% >1 mkm neighbor 24.76489
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
Peaks with higher amplitudes (> mean + 2 x SD)
original
cell#10 bottom
20 mm
10*
cell#10 bottom_part2Intensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
cell#10 bottom_part2Results
Nearest neighbors distancemean= 1.044465SD= 0.473462
# peaks total 31# >1 mkm neighbor 18% >1 mkm neighbor 58.06452
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks
Nearest neighbors distancemean= 0.709807SD= 0.22313
# peaks total 116# >1 mkm neighbor 7% >1 mkm neighbor 6.034483
Peaks with higher amplitudes (> mean + 2 x SD)
original
cell#8_top
20 mm8*
cell#8_topIntensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
Results
cell#8_top
Nearest neighbors distancemean= 0.871316SD= 0.272693
# peaks total 122# >1 mkm neighbor 35% >1 mkm neighbor 28.68852
Nearest neighbors distancemean= 1.863152SD= 1.188314
# peaks total 18# >1 mkm neighbor 11% >1 mkm neighbor 61.11111
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks Peaks with higher amplitudes (> mean + 2 x SD)
originalcell#11_bottom (main text Fig.8)
20 mm
11*
Intensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
cell#11_bottom (main text Fig.8)
Nearest neighbors distancemean= 0.90492SD= 0.277029
# peaks total 179# >1 mkm neighbor 58% >1 mkm neighbor 32.40223
Nearest neighbors distancemean= 1.272968SD= 0.925537
# peaks total 43# >1 mkm neighbor 22% >1 mkm neighbor 51.16279
Results
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks
cell#11_bottom (main text Fig.8)
Peaks with higher amplitudes (> mean + 2 x SD)
originalcell#11_top
20 mm
11*
cell#11_topIntensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
original
cell#11_topResults
Nearest neighbors distancemean= 0.871494SD= 0.320551
# peaks total 186# >1 mkm neighbor 60% >1 mkm neighbor 32.25806
Nearest neighbors distancemean= 1.229789SD= 0.691429
# peaks total 31# >1 mkm neighbor 21% >1 mkm neighbor 67.74194
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks Peaks with higher amplitudes (> mean + 2 x SD)
originalcell#13_bottom
20 mm
13*
cell#13_bottom
Intensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
cell#13_bottom
Results
Nearest neighbors distancemean= 0.7752SD= 0.275637
# peaks total 153# >1 mkm neighbor 32% >1 mkm neighbor 20.91503
Nearest neighbors distancemean= 1.122367SD= 0.528519
# peaks total 47# >1 mkm neighbor 27% >1 mkm neighbor 57.44681
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks Peaks with higher amplitudes (> mean + 2 x SD)
original cell#19_bottom
20 mm
19*
cell#19_bottomIntensity peaks of RyR labeling identified by “SD-slicing” computer algorithm (color dots).
-0.6 0 0.6 1.2 1.8 2.4 3.0 3.6 4 4.6 5.2 5.8
Peak amplitudes are coded by a rainbow-color scheme
Units are SD of total cell signalMean of total cell signal
cell#19_bottom
Results
Nearest neighbors distancemean= 1.186087SD= 1.1298
# peaks total 39# >1 mkm neighbor 20% >1 mkm neighbor 51.28205
Nearest neighbors distancemean= 0.71933SD= 0.28924
# peaks total 149# >1 mkm neighbor 21% >1 mkm neighbor 14.09396
Peak amplitude
Nearest neighbor distance
SD
mm
# of
pea
ks
# of
pea
ks
# of
pea
ks
# of
pea
ks
Nearest neighbor distance
Peak amplitude
SD
mm
All peaks Peaks with higher amplitudes (> mean + 2 x SD)