Supplementary Figure 9: Representative data analysis for STORM imaging and STORM/FLINC imaging.

(A) and (B) Double exponential fit and parameters for the spot-linking or blinking-rejection utilized in post-processing of STORM dataset in Alexa568 and Alexa647 dyes, respectively. All data were treated by spot-linking prior to further analysis to avoid over-counting. (C) Representative processing from mean-shift clustering analysis. (C1) Identification and clustering of phospho-PKA substrates localizations; (C2) Identification and clustering of AKAP79 localizations; (C3) Convex-hull cluster pairs highlighting the AKAP79 clusters (blue) and their respective phospho-PKA substrate clusters (red). (C4) Histogram displaying the frequency of occurrence for centroid distances between AKAP79 clusters and their respective nearest p-PKAsub clusters; representative of n=5 cells. (C5) Histogram displaying the frequency of occurrence for centroid distances between AKAP79 and FLINC microdomain; representative of n=5 cells. (D) Representative processing from the Getis-Franklin co-clustering analysis. (D1) Self-clustering of AKAP79 by Getis-Franklin, which is a precursor to co-clustering analysis; red color indicates increasing tendency of clustering. (D2) Co-clustering analysis plot; red color indicates increasing tendency of clustering. (D3) Representative quadrant analysis from Getis-Franklin co-clustering, reported in Figure 3C.