Extended Data Fig. 5: Processing and analysis of H. sapiens-TcmX cryo-EM structure.

a, Sorting scheme for cryo-EM data; after Initial picking, 836,588 particles were subjected to 2D-Classification, of which 461,131 particles were used for 3D-classification. Particles were sorted into four classes: low resolution particles, 80 S with E-Site, and two distinct 80 S with E-Site and eEF2. All but low-resolution particles were picked for further refinement (65.7%, 302,737 particles). After CTF-refinement and focused refinement using a 60 S mask, a final overall resolution of 2.76 Å was achieved. b, Fourier-Shell-Correlation (FSC 0.143) of the final reconstruction, with the resolution at FSC = 0.143 indicated with a dashed line. c, Overview and transverse section of cryo-EM map filtered and coloured according to local resolution. d, Electron density (mesh) and molecular model for the TcmX (blue) binding site on the human 80 S ribosome (28 S rRNA nucleotides shown in grey sticks Mg1 and Mg2 as blue spheres). e, Overview of the putative Tet-analog binding sites on the Hsa80S ribosome at the terminal loop of H89 (binding site 1, red) and within the exit tunnel (binding site 2, pink)²⁴ relative to the binding site of TcmX (blue). f, Zoom of the relative location of TcmX (blue) to 28 S rRNA nucleotides identified in binding site 2 (magenta).