Fig. 2: DNA-PAINT imaging of Nup96 in nuclear pore complex on an SDC-OPR microscope.

a Representative DNA-PAINT image of nuclear pore complexes (NPCs) in a U2OS cell, acquired using the SDC-OPR system. The imaging was performed using a DNA-labeled anti-GFP nanobody targeting Nup96, which is fluorescently tagged with mEGFP. Detailed sample preparation and imaging conditions are provided in the Methods section and Supplementary Table 1. A zoomed-in region emphasizes the resolution of individual NPCs. b Structural representation of Nup96 proteins (red) within NPCs (gray) adapted from Cryo-EM data (PDB 7PEQ). This depiction illustrates the GFP labeling of Nup96 and the corresponding DNA-labeled anti-GFP nanobody utilized in the DNA-PAINT imaging of NPCs. c Examples of individual NPCs identified in the SDC-OPR DNA-PAINT imaging, with arrows indicating specific pairs of Nup96 proteins. d Cross-sectional histogram displaying the average distances between protein pairs (n = 16) located within single symmetry centers of NPCs, highlighting the spatial organization of Nup96. Nup96 imaging was repeated in three independent experiments. Source data of histograms are provided as a Source Data file. Panel (b) was created with BioRender.com.