Particle symmetry expansion, particle subtraction and autorefinement on the CSH-tetramer + ASH monomer. (a) Generation of ACLY-E599Q–ATP–citrate–CoA (PDB 7LJW/EMD-23387) is shown as an example. (b) FSC curves illustration the improvement of ASH domain following local refinement in cryospac. The local resolution of the ASH domain was improved from 3.93.0 Å to 3.3–3.0 Å. Although the local resolution map for ACLY–OAA–acetyl-CoA (PDB 7LJ9/EMD-23389) showed a more modest improvement (4.0–2.8 Å to 3.9–2.7 Å), the map showed clearer density for the acetyl-CoA and OAA ligands (Fig. 1).