This data contains images and output data from RELION related to a publication on the mechanics of pairs of DNA-origami particles (https://www.pnas.org/doi/10.1073/pnas.2500716122). In this work, we conduct self-assembly experiments with various unique subunit types that target two different diameters of tubule structures. We provide image data of tubules that are associated with the probability distribution reported in Figure 4 in the main text for a twist-corrected monomer. Images of tubules are in the ZIP archive and show the section of tubules we analyzed to produce the probability distribution in the manuscript. This folder has an associated CSV file that relates an image name to the type of tubule that the image was identified as. Tubule types have "m" and "n" values. We provide the necessary files to recreate the fluctuation analysis that we perform on the pairs of particles. This includes output files from RELION 4’s Multi-body refinement: electron density maps of the refined bodies, a mask for the bodies, and associated data files from the principal component analysis. Accompanying these files are Python scripts for analysis. We provide results from simulations. To model the self-assembly outcomes of tubules, we perform energy minimization to find the configuration of subunits in various tubule geometries. The output of these energy minimization simulations, as well as the Python code to generate and analyze them, is provided.Associated publication citation: Thomas E.Videbæk et al., Measuring multisubunit mechanics of geometrically programmed colloidal assemblies via cryo-EM multi-body refinement, PNAS 122 (37) e2500716122 (2025). DOI:10.1073/pnas.2500716122.