Extended Data Fig. 4: Cryo-EM image processing part II (using Relion) to determine the 3D structure of RS3 in mouse respiratory cilia.
From: Mouse radial spoke 3 is a metabolic and regulatory hub in cilia
After blush refinement102 the full-length RS3 structure could be resolved, but with low resolution in the head-neck-stalk region (top right). Separate processing (for example, signal subtraction, blush refinement and 3D classification) of the head-neck and base-stalk regions improved the quality of both maps to a resolution of 7.1 Å and 4.7 Å, respectively (shown in Extended Data Fig. 5a). Each step uses the results of the preceding step as a direct starting point, including references for 3D refinement and classification. The composite map of the full-length RS3 was generated by aligning the two maps based on their overlapping region in ChimeraX100.
