Cryo-EM 2D class averages of MST1 and cryo-EM flowchart. (A) Representative 2D class averages obtained from cryo-images are displayed with inverted contrast. (B) Cryo-EM workflow for mastigoneme (further details provided in the Materials and methods section). The flowchart includes FSC (Fourier Shell Correlation) curves with a gold standard threshold of 0.143 for mastigoneme. Additionally, the angular distribution, the particle distribution plot, original maps, and 3D FSC analysis of mastigoneme are shown. The local resolution analysis of representative cryo-EM densities for mastigoneme is also presented.