Fig. 4: Clustering of RyR2 subunit localizations in HEK293 cells.

A Overview of RyR2 subunit clustering in a HEK293 cell expressing RyR2_D4365-GFP. Clustering was determined with DBSCAN (ε = 100 nm) and using the 3D coordinates of subunit locations. Coloring indicates cluster size. B x-z view of the large yellow colored cluster in (A). C, D. x-z and y-z views of the loop-region indicated in B. Coloring in B-D indicates z elevation. E Boxplots of mean cluster sizes measured in SU/cluster as counted from data (Measured) and after correction with an indicative effective labeling efficiency (Corrected). N = 10 MINFLUX data sets from n = 3 biological replicates. F Fractions of RyRs present in the three largest cluster indicating that a sizable fraction and frequently the majority of RyRs are within a few giant clusters. N = 10 MINFLUX data sets from n = 3 biological replicates. G Mean cluster sizes across all data sets obtained by using labeling efficiency corrected data from E and division by 4 to obtain RyR2 numbers from SU numbers. N = 10 MINFLUX data sets from n = 3 biological replicates. Box plots show the median (center line), the 25th and 75th percentiles (bounds of the box), and the mean (large circular marker). Whiskers extend to the most extreme data points within 1.5x the interquartile range, and all individual data points are displayed as a swarm overlay. Scale bars A: 1 µm; B: 500 nm; C, D: 250 nm. Source data are provided with this paper.