Extended Data Fig. 2: Workflow for negative-stain EM data processing.
From: Nde1 promotes Lis1 binding to full-length autoinhibited human dynein 1
a, Representative micrographs for dynein alone (42 micrographs), dynein-Lis1 at 1:1 (41 micrographs), and dynein-Lis1 at 1:2 (47 micrographs) molar ratios from batch #1. b, Particle picking from representative micrographs in each dataset using a template matching approach based on Phi and open dynein models (particle diameter: 750 Å, distance cutoff: 400 Å). c, Three rounds of 2D classification were performed after extracting all particles (box size: 960 Å × 960 Å), yielding class averages of Phi dynein, open dynein motors, single motors, and junk particles. d, Final classified 2D averages showing Phi dynein (43.7%), two-motor open dynein (43.3%), and single-motor open dynein (13.0%). The particle numbers for each group were counted, and single motors were considered as open dynein by dividing the total number of particles by two.
