Counting the number of Rtn4 copies per oligomer. (A) Representative whole field-of-view of a dataset. A U-2 OS Rtn4-mEGFP cell and a U-2 OS Nup96-mEGFP cell are located in the bottom left and the upper right of the image, respectively. (B) Zoom-in of Nup96-mEGFP. (C) Histograms from each experiment of the number of localizations per nuclear pore complex. (D) Zoom-in of Rtn4-mEGFP. (E) Same zoom-in as D after segmenting clusters of Rtn4 localizations using DBSCAN. Clusters are colored uniquely. (F) Histograms from each experiment (experiment #1: blue, experiment #2: orange, experiment #3: green) of the number of Rtn4 copies per cluster of Rtn4 localizations. Median values of Rtn4 copies per cluster—experiment #1: 5.36, experiment #2: 5.67, experiment #3: 4.75. Inset shows the same plot with the x-axis limited to 10 copies of Rtn4 per cluster. Scale bars represent 1 μm.