Cryo-EM analysis of α-HL arc-like and octameric species in prehemolytic RBCs. (A) Cryo-EM data processing pipeline of α-HL arc-like species. Different sizes of arcs are highlighted in different square boxes (pink—dimer; blue—4-mer; white—5-mer; and orange—6-mer). (B) 3D refinement of the pentameric/5-mer species, and model fitting different protomers shown in different colors in the EM map (gray color). The prestem detached protomer was used from our pore structure, and for the prestem attached monomer (blue color), the PDB ID: 4YHD structure was used. (C) Orientation of the pentameric structure. The red-box region shows the zoomed view of the N-terminal region. In the zoomed view, the N terminus of one of the protomers is highlighted in golden rod color, showing its interaction with the upper cap domain. (D) Cryo-EM data processing pipeline of α-HL octamers in prehemolytic RBCs.