NS-TEM and cryo-EM analysis of prehemolytic and posthemolytic states of RBCs. (A) NS-TEM images of control RBCs (low-magnification image at the left side and high-magnification image at the right side). (B) NS-TEM images of toxin–RBCs (low-magnification image at the left side and high-magnification image at the right side). 30 images were collected from different regions in triplicate to validate this phenomenon. (C) Cryo-EM image of control RBC (entire cell). (D) Cryo-EM image of RBC (entire cell) in prehemolytic condition and its zoomed image. (E) Cryo-EM image of RBC (entire cell) in posthemolytic condition and its zoomed view. (F and G) Cryo-EM micrographs of toxin-containing membranes, either in lysed form or in vesicular form, outside the RBCs in post- and prehemolytic conditions, respectively (scale bar: 100 nm).